US 2004/0167066 A1Published application

Cleavage and polyadenylation complex of precursor mrna

OwnerMartina Marzioch, Anne-Claude Gavin +4inventors

InventorsMartina Marzioch, Anne-Claude Gavin, Andreas Bauer, Jorg Schultz, Miro Brajenovic, Paola Grandi

Life of this patent

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Abstract

The present invention relates to novel components of the cleavage/polyadenylation machinery of precursor mRNA as well as to the complex containing the new components and its use. The complex is obtained by using one component thereof as a bait and isolating a highly organized complex consisting of at least 13 distinct proteins.

Claims45

Dependent claims

An isolated complex selected from complex (I) and comprising

(a) a first protein, or a functionally active fragment or functionally active derivative thereof, which first protein is selected from the group consisting of:
(i) Cft1 (SEQ ID NO:3), or a mammalian homolog thereof, or a variant of Cft1 encoded by a nucleic acid that hybridizes to the Cft1 nucleic acid (SEQ ID NO:4) or its complement under low stringency conditions,
(ii) Cft2 (SEQ ID NO:5), or a mammalian homolog thereof, or a variant of Cft2 encoded by a nucleic acid that hybridizes to the Cft2 nucleic acid (SEQ ID NO:6) or its complement under low stringency conditions,
(iii) Clp1 (SEQ ID NO:9), or a mammalian homolog thereof, or a variant of Clp1 encoded by a nucleic acid that hybridizes to the Clp1 nucleic acid (SEQ ID NO:10) or its complement under low stringency conditions,
(iv) Fip1 (SEQ ID NO:19), or a mammalian homolog thereof, or a variant of Fip1 encoded by a nucleic acid that hybridizes to the Fip1 nucleic acid (SEQ ID NO:20) or its complement under low stringency conditions,
(v) Pab1 (SEQ ID NO:33), or a mammalian homolog thereof, or a variant of Pab1 encoded by a nucleic acid that hybridizes to the Pab1 nucleic acid (SEQ ID NO:34) or its complement under low stringency conditions,
(vi) Pap1 (SEQ ID NO:35), or a mammalian homolog thereof, or a variant of Pap1 encoded by a nucleic acid that hybridizes to the Pap1 nucleic acid (SEQ ID NO:36) or its complement under low stringency conditions,
(vii) Pcf11 (SEQ ID NO:37), or a mammalian homolog thereof, or a variant of Pcf11 encoded by a nucleic acid that hybridizes to the Pcf11 nucleic acid (SEQ ID NO:38) or its complement under low stringency conditions,
(viii) Pfs2 (SEQ ID NO:43), or a mammalian homolog thereof, or a variant of Pfs2 encoded by a nucleic acid that hybridizes to the Pfs2 nucleic acid (SEQ ID NO:44) or its complement under low stringency conditions,
(ix) Pta1 (SEQ ID NO:45), or a mammalian homolog thereof, or a variant of Pta1 encoded by a nucleic acid that hybridizes to the Pta1 nucleic acid (SEQ ID NO:46) or its complement under low stringency conditions,
(x) Rna14 (SEQ ID NO:49), or a mammalian homolog thereof, or a variant of Rna14 encoded by a nucleic acid that hybridizes to the Rna14 nucleic acid (SEQ ID NO:50) or its complement under low stringency conditions,
(xi) Rna15 (SEQ ID NO:51), or a mammalian homolog thereof, or a variant of Rna15 encoded by a nucleic acid that hybridizes to the Rna15 nucleic acid (SEQ ID NO:52) or its complement under low stringency conditions,
(xii) Tif4632 (SEQ ID NO:63), or a mammalian homolog thereof, or a variant of Tif4632 encoded by a nucleic acid that hybridizes to the Tif4632 nucleic acid (SEQ ID NO:64) or its complement under low stringency conditions,
(xiii) Ykl059c (SEQ ID NO:89), or a mammalian homolog thereof, or a variant of Ykl059c encoded by a nucleic acid that hybridizes to the Ykl059c nucleic acid (SEQ ID NO:90) or its complement under low stringency conditions,
(xiv) Ysh1 (SEQ ID NO:75), or a mammalian homolog thereof, or a variant of Ysh1 encoded by a nucleic acid that hybridizes to the Ysh1 nucleic acid (SEQ ID NO:76) or its complement under low stringency conditions, and
(xv) Yth1 (SEQ ID NO:77), or a mammalian homolog thereof, or a variant of Yth1 encoded by a nucleic acid that hybridizes to the Yth1 nucleic acid (SEQ ID NO:78) or its complement under low stringency conditions;
and
(b) a second protein, or a functionally active fragment or functionally active derivative thereof, which second protein is selected from the group consisting of:
(i) Act1 (SEQ ID NO:1), or a mammalian homolog thereof, or a variant of Act1 encoded by a nucleic acid that hybridizes to the Act1 nucleic acid (SEQ ID NO:2) or its complement under low stringency conditions,
(ii) Cka1 (SEQ ID NO:7), or a mammalian homolog thereof, or a variant of Cka1 encoded by a nucleic acid that hybridizes to the Cka1 nucleic acid (SEQ ID NO:8) or its complement under low stringency conditions,
(iii) Eft2 (SEQ ID NO:11), or a mammalian homolog thereof, or a variant of Eft2 encoded by a nucleic acid that hybridizes to the Eft2 nucleic acid (SEQ ID NO:12) or its complement under low stringency conditions,
(iv) Eno2 (SEQ ID NO:13), or a mammalian homolog thereof, or a variant of Eno2 encoded by a nucleic acid that hybridizes to the Eno2 nucleic acid (SEQ ID NO:14) or its complement under low stringency conditions,
(v) Glc7 (SEQ ID NO:15), or a mammalian homolog thereof, or a variant of Glc7 encoded by a nucleic acid that hybridizes to the Glc7 nucleic acid (SEQ ID NO:16) or its complement under low stringency conditions,
(vi) Gpm1 (SEQ ID NO:17), or a mammalian homolog thereof, or a variant of Gpm1 encoded by a nucleic acid that hybridizes to the Gpm1 nucleic acid (SEQ ID NO:18) or its complement under low stringency conditions,
(vii) Hhf2 (SEQ ID NO:21), or a mammalian homolog thereof, or a variant of Hhf2 encoded by a nucleic acid that hybridizes to the Hhf2 nucleic acid (SEQ ID NO:22) or its complement under low stringency conditions,
(viii) Hta1 (SEQ ID NO:23), or a mammalian homolog thereof, or a variant of Hta1 encoded by a nucleic acid that hybridizes to the Hta1 nucleic acid (SEQ ID NO:24) or its complement under low stringency conditions,
(ix) Hsc82 (SEQ ID NO:25), or a mammalian homolog thereof, or a variant of Hsc82 encoded by a nucleic acid that hybridizes to the Hsc82 nucleic acid (SEQ ID NO:26) or its complement under low stringency conditions,
(x) Imd2 (SEQ ID NO:27), or a mammalian homolog thereof, or a variant of Imd2 encoded by a nucleic acid that hybridizes to the Imd2 nucleic acid (SEQ ID NO:28) or its complement under low stringency conditions,
(xi) Imd4 (SEQ ID NO:29), or a mammalian homolog thereof, or a variant of Imd4 encoded by a nucleic acid that hybridizes to the Imd4 nucleic acid (SEQ ID NO:30) or its complement under low stringency conditions,
(xii) Met6 (SEQ ID NO:31), or a mammalian homolog thereof, or a variant of Met6 encoded by a nucleic acid that hybridizes to the Met6 nucleic acid (SEQ ID NO:32) or its complement under low stringency conditions,
(xiii) Pdc1 (SEQ ID NO:39), or a mammalian homolog thereof, or a variant of Pdc1 encoded by a nucleic acid that hybridizes to the Pdc1 nucleic acid (SEQ ID NO:40) or its complement under low stringency conditions,
(xiv) Pfk1 (SEQ ID NO:41), or a mammalian homolog thereof, or a variant of Pfk1 encoded by a nucleic acid that hybridizes to the Pfk1 nucleic acid (SEQ ID NO:42) or its complement under low stringency conditions,
(xv) Ref2 (SEQ ID NO:47), or a mammalian homolog thereof, or a variant of Ref2 encoded by a nucleic acid that hybridizes to the Ref2 nucleic acid (SEQ ID NO:48) or its complement under low stringency conditions,
(xvi) Sec13 (SEQ ID NO:53), or a mammalian homolog thereof, or a variant of Sec13 encoded by a nucleic acid that hybridizes to the Sec13 nucleic acid (SEQ ID NO:54) or its complement under low stringency conditions,
(xvii) Sec31 (SEQ ID NO:55), or a mammalian homolog thereof, or a variant of Sec31 encoded by a nucleic acid that hybridizes to the Sec31 nucleic acid (SEQ ID NO:56) or its complement under low stringency conditions,
(xviii) Ssa3 (SEQ ID NO:57), or a mammalian homolog thereof, or a variant of Ssa3 encoded by a nucleic acid that hybridizes to the Ssa3 nucleic acid (SEQ ID NO:58) or its complement under low stringency conditions,
(xix) Ssu72 (SEQ ID NO:59), or a mammalian homolog thereof, or a variant of Ssu72 encoded by a nucleic acid that hybridizes to the Ssu72 nucleic acid (SEQ ID NO:60) or its complement under low stringency conditions,
(xx) Taf60 (SEQ ID NO:61), or a mammalian homolog thereof, or a variant of Taf60 encoded by a nucleic acid that hybridizes to the Taf60 nucleic acid (SEQ ID NO:62) or its complement under low stringency conditions,
(xxi) Tkl1 (SEQ ID NO:65), or a mammalian homolog thereof, or a variant of Tkl1 encoded by a nucleic acid that hybridizes to the Tkl1 nucleic acid (SEQ ID NO:66) or its complement under low stringency conditions,
(xxii) Tsa1 (SEQ ID NO:67), or a mammalian homolog thereof, or a variant of Tsa1 encoded by a nucleic acid that hybridizes to the Tsa1 nucleic acid (SEQ ID NO:68) or its complement under low stringency conditions,
(xxiii) Tye7 (SEQ ID NO:69), or a mammalian homolog thereof, or a variant of Tye7 encoded by a nucleic acid that hybridizes to the Tye7 nucleic acid (SEQ ID NO:70) or its complement under low stringency conditions,
(xxiv) Vid24 (SEQ ID NO:71), or a mammalian homolog thereof, or a variant of Vid24 encoded by a nucleic acid that hybridizes to the Vid24 nucleic acid (SEQ ID NO:72) or its complement under low stringency conditions,
(xxv) Vps53 (SEQ ID NO:73), or a mammalian homolog thereof, or a variant of Vps53 encoded by a nucleic acid that hybridizes to the Vps53 nucleic acid (SEQ ID NO:74) or its complement under low stringency conditions,
(xxvi) Ycl046w (SEQ ID NO:79), or a mammalian homolog thereof, or a variant of Ycl046w encoded by a nucleic acid that hybridizes to the Ycl046w nucleic acid (SEQ ID NO:80) or its complement under low stringency conditions,
(xxvii) Ygr156w (SEQ ID NO:81), or a mammalian homolog thereof, or a variant of Ygr156w encoded by a nucleic acid that hybridizes to the Ygr156w nucleic acid (SEQ ID NO:82) or its complement under low stringency conditions,
(xxviii) Yhl035c (SEQ ID NO:83), or a mammalian homolog thereof, or a variant of Yhl035c encoded by a nucleic acid that hybridizes to the Yhl035c nucleic acid (SEQ ID NO:84) or-its complement under low stringency conditions,
(xxix) Ykl018w (SEQ ID NO:85), or a mammalian homolog thereof, or a variant of Ykl018w encoded by a nucleic acid that hybridizes to the Ykl018w nucleic acid (SEQ ID NO:86) or its complement under low stringency conditions,
(xxx) Ylr221c (SEQ ID NO:87), or a mammalian homolog thereof, or a variant of Ylr221c encoded by a nucleic acid that hybridizes to the Ylr221c nucleic acid (SEQ ID NO:88) or its complement under low stringency conditions,
(xxxi) Yml030w (SEQ ID NO:91), or a mammalian homolog thereof, or a variant of Yml030w encoded by a nucleic acid that hybridizes to the Yml030w nucleic acid (SEQ ID NO:92) or its complement under low stringency conditions, and
(xxxii) Yor179c (SEQ ID NO:93), or a mammalian homolog thereof, or a variant of Yor179c encoded by a nucleic acid that hybridizes to the Yor179c nucleic acid (SEQ ID NO:94) or its complement under low stringency conditions,
wherein said first protein and said second protein are members of a native cellular Polyadenylation-complex, and wherein said low stringency conditions comprise hybridization in a buffer comprising 35% formamide, 5×SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 ug/ml denatured salmon sperm DNA, and 10% (wt/vol) dextran sulfate for 18-20 hours at 40° C., washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 55° C., and washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 60° C., and a complex (II) comprising at least two second proteins.

48 more elements

An isolated complex comprising the following proteins:

(i) Act1 (SEQ ID NO:1), or a mammalian homolog thereof, or a variant of Act1 encoded by a nucleic acid that hybridizes to the Act1 nucleic acid (SEQ ID NO:2) or its complement under low stringency conditions,
(ii) Cft1 (SEQ ID NO:3), or a mammalian homolog thereof, or a variant of Cft1 encoded by a nucleic acid that hybridizes to the Cft1 nucleic acid (SEQ ID NO:4) or its complement under low stringency conditions,
(iii) Cft2 (SEQ ID NO:5), or a mammalian homolog thereof, or a variant of Cft2 encoded by a nucleic acid that hybridizes to the Cft2 nucleic acid (SEQ ID NO:6) or its complement under low stringency conditions,
(iv) Cka1 (SEQ ID NO:7), or a mammalian homolog thereof, or a variant of Cka1 encoded by a nucleic acid that hybridizes to the Cka1 nucleic acid (SEQ ID NO:8) or its complement under low stringency conditions,
(v) Clp1 (SEQ ID NO:9), or a mammalian homolog thereof, or a variant of Clp1 encoded by a nucleic acid that hybridizes to the Clp1 nucleic acid (SEQ ID NO:10) or its complement under low stringency conditions,
(vi) Eft2 (SEQ ID NO:11), or a mammalian homolog thereof, or a variant of Eft2 encoded by a nucleic acid that hybridizes to the Eft2 nucleic acid (SEQ ID NO:12) or its complement under low stringency conditions,
(vii) Eno2 (SEQ ID NO:13), or a mammalian homolog thereof, or a variant of Eno2 encoded by a nucleic acid that hybridizes to the Eno2 nucleic acid (SEQ ID NO:14) or its complement under low stringency conditions,
(viii) Glc7 (SEQ ID NO:15), or a mammalian homolog thereof, or a variant of Glc7 encoded by a nucleic acid that hybridizes to the Glc7 nucleic acid (SEQ ID NO:16) or its complement under low stringency conditions,
(ix) Gpm1 (SEQ ID NO:17), or a mammalian homolog thereof, or a variant of Gpm1 encoded by a nucleic acid that hybridizes to the Gpm1 nucleic acid (SEQ ID NO:18) or its complement under low stringency conditions,
(x) Fip1 (SEQ ID NO:19), or a mammalian homolog thereof, or a variant of Fip1 encoded by a nucleic acid that hybridizes to the Fip1 nucleic acid (SEQ ID NO:20) or its complement under low stringency conditions,
(xi) Hhf2 (SEQ ID NO:21), or a mammalian homolog thereof, or a variant of Hhf2 encoded by a nucleic acid that hybridizes to the Hhf2 nucleic acid (SEQ ID NO:22) or its complement under low stringency conditions,
(xii) Hta1 (SEQ ID NO:23), or a mammalian homolog thereof, or a variant of Hta1 encoded by a nucleic acid that hybridizes to the Hta1 nucleic acid (SEQ ID NO:24) or its complement under low stringency conditions,
(xiii) Hsc82 (SEQ ID NO:25), or a mammalian homolog thereof, or a variant of Hsc82 encoded by a nucleic acid that hybridizes to the Hsc82 nucleic acid (SEQ ID NO:26) or its complement under low stringency conditions,
(xiv) Imd2 (SEQ ID NO:27), or a mammalian homolog thereof, or a variant of Imd2 encoded by a nucleic acid that hybridizes to the Imd2 nucleic acid (SEQ ID NO:28) or its complement under low stringency conditions,
(xv) Imd4 (SEQ ID NO:29), or a mammalian homolog thereof, or a variant of Imd4 encoded by a nucleic acid that hybridizes to the Imd4 nucleic acid (SEQ ID NO:30) or its complement under low stringency conditions,
(xvi) Met6 (SEQ ID NO:31), or a mammalian homolog thereof, or a variant of Met6 encoded by a nucleic acid that hybridizes to the Met6 nucleic acid (SEQ ID NO:32) or its complement under low stringency conditions,
(xvii) Pab1 (SEQ ID NO:33), or a mammalian homolog thereof, or a variant of Pab1 encoded by a nucleic acid that hybridizes to the Pab1 nucleic acid (SEQ ID NO:34) or its complement under low stringency conditions,
(xviii) Pap1 (SEQ ID NO:35), or a mammalian homolog thereof, or a variant of Pap1 encoded by a nucleic acid that hybridizes to the Pap1 nucleic acid (SEQ ID NO:36) or its complement under low stringency conditions,
(xix) Pcf11 (SEQ ID NO:37), or a mammalian homolog thereof, or a variant of Pcf11 encoded by a nucleic acid that hybridizes to the Pcf11 nucleic acid (SEQ ID NO:38) or its complement under low stringency conditions,
(xx) Pdc1 (SEQ ID NO:39), or a mammalian homolog thereof, or a variant of Pdc1 encoded by a nucleic acid that hybridizes to the Pdc1 nucleic acid (SEQ ID NO:40) or its complement under low stringency conditions,
(xxi) Pfk1 (SEQ ID NO:41), or a mammalian homolog thereof, or a variant of Pfk1 encoded by a nucleic acid that hybridizes to the Pfk1 nucleic acid (SEQ ID NO:42) or its complement under low stringency conditions,
(xxii) Pfs2 (SEQ ID NO:43), or a mammalian homolog thereof, or a variant of Pfs2 encoded by a nucleic acid that hybridizes to the Pfs2 nucleic acid (SEQ ID NO:44) or its complement under low stringency conditions,
(xxiii) Pta1 (SEQ ID NO:45), or a mammalian homolog thereof, or a variant of Pta1 encoded by a nucleic acid that hybridizes to the Pta1 nucleic acid (SEQ ID NO:46) or its complement under low stringency conditions,
(xxiv) Ref2 (SEQ ID NO:47), or a mammalian homolog thereof, or a variant of Ref2 encoded by a nucleic acid that hybridizes to the Ref2 nucleic acid (SEQ ID NO:48) or its complement under low stringency conditions,
(xxv) Rna14 (SEQ ID NO:49), or a mammalian homolog thereof, or a variant of Rna14 encoded by a nucleic acid that hybridizes to the Rna14 nucleic acid (SEQ ID NO:50) or its complement under low stringency conditions,
(xxvi) Rna15 (SEQ ID NO:51), or a mammalian homolog thereof, or a variant of Rna15 encoded by a nucleic acid that hybridizes to the Rna15 nucleic acid (SEQ ID NO:52) or its complement under low stringency conditions,
(xxvii) Sec13 (SEQ ID NO:53), or a mammalian homolog thereof, or a variant of Sec13 encoded by a nucleic acid that hybridizes to the Sec13 nucleic acid (SEQ ID NO:54) or its complement under low stringency conditions,
(xxviii) Sec31 (SEQ ID NO:55), or a mammalian homolog thereof, or a variant of Ser31 encoded by a nucleic acid that hybridizes to the Sec31 nucleic acid (SEQ ID NO:56) or its complement under low stringency conditions,
(xxix) Ssa3 (SEQ ID NO:57), or a mammalian homolog thereof, or a variant of Ssa3 encoded by a nucleic acid that hybridizes to the Ssa3 nucleic acid (SEQ ID NO:58) or its complement under low stringency conditions,
(xxx) Ssu72 (SEQ ID NO:59), or a mammalian homolog thereof, or a variant of Ssu72 encoded by a nucleic acid that hybridizes to the Ssu72 nucleic acid (SEQ ID NO:60) or its complement under low stringency conditions,
(xxxi) Taf60 (SEQ ID NO:61), or a mammalian homolog thereof, or a variant of Taf60 encoded by a nucleic acid that hybridizes to the Taf60 nucleic acid (SEQ ID NO:62) or its complement under low stringency conditions,
(xxxii) Tif4632 (SEQ ID NO:63), or a mammalian homolog thereof, or a variant of Tif4632 encoded by a nucleic acid that hybridizes to the Tif4632 nucleic acid (SEQ ID NO:64) or its complement under low stringency conditions,
(xxxiii) Tkl1 (SEQ ID NO:65), or a mammalian homolog thereof, or a variant of Tkl1 encoded by a nucleic acid that hybridizes to the Tkl1 nucleic acid (SEQ ID NO:66) or its complement under low stringency conditions,
(xxxiv) Tsa1 (SEQ ID NO:67), or a mammalian homolog thereof, or a variant of Tsa1 encoded by a nucleic acid that hybridizes to the Tsa1 nucleic acid (SEQ ID NO:68) or its complement under low stringency conditions,
(xxxv) Tye7 (SEQ ID NO:69), or a mammalian homolog thereof, or a variant of Tye7 encoded by a nucleic acid that hybridizes to the Tye7 nucleic acid (SEQ ID NO:70) or its complement under low stringency conditions,
(xxxvi) Vid24 (SEQ ID NO:71), or a mammalian homolog thereof, or a variant of Vid24 encoded by a nucleic acid that hybridizes to the Vid24 nucleic acid (SEQ ID NO:72) or its complement under low stringency conditions,
(xxxvii) Vps53 (SEQ ID NO:73), or a mammalian homolog thereof, or a variant of Vps53 encoded by a nucleic acid that hybridizes to the Vps53 nucleic acid (SEQ ID NO:74) or its complement under low stringency conditions,
(xxxviii) Ysh1 (SEQ ID NO:75), or a mammalian homolog thereof, or a variant of Ysh1 encoded by a nucleic acid that hybridizes to the Ysh1 nucleic acid (SEQ ID NO:76) or its complement under low stringency conditions,
(xxxix) Yth1 (SEQ ID NO:77), or a mammalian homolog thereof, or a variant of Yth1 encoded by a nucleic acid that hybridizes to the Yth1 nucleic acid (SEQ ID NO:78) or its complement under low stringency conditions,
(xl) Ycl046w (SEQ ID NO:79), or a mammalian homolog thereof, or a variant of Ycl046w encoded by a nucleic acid that hybridizes to the Ycl046w nucleic acid (SEQ ID NO:80) or its complement under low stringency conditions,
(xli) Ygr156w (SEQ ID NO:81), or a mammalian homolog thereof, or a variant of Ygr156w encoded by a nucleic acid that hybridizes to the Ygr156w nucleic acid (SEQ ID NO:82) or its complement under low stringency conditions,
(xlii) Yhl035c (SEQ ID NO:83), or a mammalian homolog thereof, or a variant of Yhl035c encoded by a nucleic acid that hybridizes to the Yhl035c nucleic acid (SEQ ID NO:84) or its complement under low stringency conditions,
(xliii) Ykl018w (SEQ ID NO:85), or a mammalian homolog thereof, or a variant of Ykl018w encoded by a nucleic acid that hybridizes to the Ykl018w nucleic acid (SEQ ID NO:86) or its complement under low stringency conditions,
(xliv) Ylr221c (SEQ ID NO:87), or a mammalian homolog thereof, or a variant of Ylr221c encoded by a nucleic acid that hybridizes to the Ylr221c nucleic acid (SEQ ID NO:88) or its complement under low stringency conditions,
(xlv) Ykl059c (SEQ ID NO:89), or a mammalian homolog thereof, or a variant of Ykl059c encoded by a nucleic acid that hybridizes to the Ykl059c nucleic acid (SEQ ID NO:90) or its complement under low stringency conditions,
(xlvi) Yml030w (SEQ ID NO:91), or a mammalian homolog thereof, or a variant of Yml030w encoded by a nucleic acid that hybridizes to the Yml030w nucleic acid (SEQ ID NO:92) or its complement under low stringency conditions, and
(xlvii) Yor179c (SEQ ID NO:93), or a mammalian homolog thereof, or a variant of Yor179c encoded by a nucleic acid that hybridizes to the Yor179c nucleic acid (SEQ ID NO:94) or its complement under low stringency conditions,
wherein said proteins are members of a native cellular Polyadenylation-complex, and
wherein said low stringency conditions comprise hybridization in a buffer comprising 35% formamide, 5×SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 ug/ml denatured salmon sperm DNA, and 10% (wt/vol) dextran sulfate for 18-20 hours at 40° C., washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 55° C., and washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 60° C.

46 more elements

An isolated complex that comprises all but 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18,19,20,21,22,23,24,25,26,27 or 28 of the following 47 proteins:

(i) Act1 (SEQ ID NO:1), or a mammalian homolog thereof, or a variant of Act1 encoded by a nucleic acid that hybridizes to the Act1 nucleic acid (SEQ ID NO:2) or its complement under low stringency conditions,
(ii) Cft1 (SEQ ID NO:3), or a mammalian homolog thereof, or a variant of Cft1 encoded by a nucleic acid that hybridizes to the Cft1 nucleic acid (SEQ ID NO-4) or its complement under low stringency conditions,
(iii) Cft2 (SEQ ID NO:5), or a mammalian homolog thereof, or a variant of Cft2 encoded by a nucleic acid that hybridizes to the Cft2 nucleic acid (SEQ ID NO:6) or its complement under low stringency conditions,
(iv) Cka1 (SEQ ID NO:7), or a mammalian homolog thereof, or a variant of Cka1 encoded by a nucleic acid that hybridizes to the Cka1 nucleic acid (SEQ ID NO:8) or its complement under low stringency conditions,
(v) Clp1 (SEQ ID NO:9), or a mammalian homolog thereof, or a variant of Clp1 encoded by a nucleic acid that hybridizes to the Clp1 nucleic acid (SEQ ID NO:10) or its complement under low stringency conditions,
(vi) Eft2 (SEQ ID NO:11), or a mammalian homolog thereof, or a variant of Eft2 encoded by a nucleic acid that hybridizes to the Eft2 nucleic acid (SEQ ID NO:12) or its complement under low stringency conditions,
(vii) Eno2 (SEQ ID NO:13), or a mammalian homolog thereof, or a variant of Eno2 encoded by a nucleic acid that hybridizes to the Eno2 nucleic acid (SEQ ID NO:14) or its complement under low stringency conditions,
(viii) Glc7 (SEQ ID NO:15), or a mammalian homolog thereof, or a variant of Glc7 encoded by a nucleic acid that hybridizes to the Glc7 nucleic acid (SEQ ID NO:16) or its complement under low stringency conditions,
(ix) Gpm1 (SEQ ID NO:17), or a mammalian homolog thereof, or a variant of Gpm1 encoded by a nucleic acid that hybridizes to the Gpm1 nucleic acid (SEQ ID NO:18) or its complement under low stringency conditions,
(x) Fip1 (SEQ ID NO:19), or a mammalian homolog thereof, or a variant of Fip1 encoded by a nucleic acid that hybridizes to the Fip1 nucleic acid (SEQ ID NO:20) or its complement under low stringency conditions,
(xi) Hhf2 (SEQ ID NO:21), or a mammalian homolog thereof, or a variant of Hhf2 encoded by a nucleic acid that hybridizes to the Hhf2 nucleic acid (SEQ ID NO:22) or its complement under low stringency conditions,
(xii) Hta1 (SEQ ID NO:23), or a mammalian homolog thereof, or a variant of Hta1 encoded by a nucleic acid that hybridizes to the Hta1 nucleic acid (SEQ ID NO:24) or its complement under low stringency conditions,
(xiii) Hsc82 (SEQ ID NO:25), or a mammalian homolog thereof, or a variant of Hsc82 encoded by a nucleic acid that hybridizes to the Hsc82 nucleic acid (SEQ ID NO:26) or its complement under low stringency conditions,
(xiv) Imd2 (SEQ ID NO:27), or a mammalian homolog thereof, or a variant of Imd2 encoded by a nucleic acid that hybridizes to the Imd2 nucleic acid (SEQ ID NO:28) or its complement under low stringency conditions,
(xv) Imd4 (SEQ ID NO:29), or a mammalian homolog thereof, or a variant of Imd4 encoded by a nucleic acid that hybridizes to the Imd4 nucleic acid (SEQ ID NO:30) or its complement under low stringency conditions,
(xvi) Met6 (SEQ ID NO:31), or a mammalian homolog thereof, or a variant of Met6 encoded by a nucleic acid that hybridizes to the Met6 nucleic acid (SEQ ID NO:32) or its complement under low stringency conditions,
(xvii) Pab1 (SEQ ID NO:33), or a mammalian homolog thereof, or a variant of Pab1 encoded by a nucleic acid that hybridizes to the Pab1 nucleic acid (SEQ ID NO:34) or its complement under low stringency conditions,
(xviii) Pap1 (SEQ ID NO:35), or a mammalian homolog thereof, or a variant of Pap1 encoded by a nucleic acid that hybridizes to the Pap1 nucleic acid (SEQ ID NO:36) or its complement under low stringency conditions,
(xix) Pcf11 (SEQ ID NO:37), or a mammalian homolog thereof, or a variant of Pcf11 encoded by a nucleic acid that hybridizes to the Pcf11 nucleic acid (SEQ ID NO:38) or its complement under low stringency conditions,
(xx) Pdc1 (SEQ ID NO:39), or a mammalian homolog thereof, or a variant of Pdc1 encoded by a nucleic acid that hybridizes to the Pdc1 nucleic acid (SEQ ID NO:40) or its complement under low stringency conditions,
(xxi) Pfk1 (SEQ ID NO:41), or a mammalian homolog thereof, or a variant of Pfk1 encoded by a nucleic acid that hybridizes to the Pfk1 nucleic acid (SEQ ID NO:42) or its complement under low stringency conditions,
(xxii) Pfs2 (SEQ ID NO:43), or a mammalian homolog thereof, or a variant of Pfs2 encoded by a nucleic acid that hybridizes to the Pfs2 nucleic acid (SEQ ID NO:44) or its complement under low stringency conditions,
(xxiii) Pta1 (SEQ ID NO:45), or a mammalian homolog thereof, or a variant of Pta1 encoded by a nucleic acid that hybridizes to the Pta1 nucleic acid (SEQ ID NO:46) or its complement under low stringency conditions,
(xxiv) Ref2 (SEQ ID NO:47), or a mammalian homolog thereof, or a variant of Ref2 encoded by a nucleic acid that hybridizes to the Ref2 nucleic acid (SEQ ID NO:48) or its complement under low stringency conditions,
(xxv) Rna14 (SEQ ID NO:49), or a mammalian homolog thereof, or a variant of Rna14 encoded by a nucleic acid that hybridizes to the Rna14 nucleic acid (SEQ ID NO:50) or its complement under low stringency conditions,
(xxvi) Rna15 (SEQ ID NO:51), or a mammalian homolog thereof, or a variant of Rna15 encoded by a nucleic acid that hybridizes to the Rna15 nucleic acid (SEQ ID NO:52) or its complement under low stringency conditions,
(xxvii) Sec13 (SEQ ID NO:53), or a mammalian homolog thereof, or a variant of Sec13 encoded by a nucleic acid that hybridizes to the Sec13 nucleic acid (SEQ ID NO:54) or its complement under low stringency conditions,
(xxviii) Sec31 (SEQ ID NO:55), or a mammalian homolog thereof, or a variant of Sec31 encoded by a nucleic acid that hybridizes to the Sec31 nucleic acid (SEQ ID NO:56) or its complement under low stringency conditions,
(xxix) Ssa3 (SEQ ID NO:57), or a mammalian homolog thereof, or a variant of Ssa3 encoded by a nucleic acid that hybridizes to the Ssa3 nucleic acid (SEQ ID NO:58) or its complement under low stringency conditions,
(xxx) Ssu72 (SEQ ID NO:59), or a mammalian homolog thereof, or a variant of Ssu72 encoded by a nucleic acid that hybridizes to the Ssu72 nucleic acid (SEQ ID NO:60) or its complement under low stringency conditions,
(xxxi) Taf60 (SEQ ID NO:61), or a mammalian homolog thereof, or a variant of Taf60 encoded by a nucleic acid that hybridizes to the Taf60 nucleic acid (SEQ ID NO:62) or its complement under low stringency conditions,
(xxxii) Tif4632 (SEQ ID NO:63), or a mammalian homolog thereof, or a variant of Tif4632 encoded by a nucleic acid that hybridizes to the Tif4632 nucleic acid (SEQ ID NO:64) or its complement under low stringency conditions,
(xxxiii) Tkl1 (SEQ ID NO:65), or a mammalian homolog thereof, or a variant of Tkl1 encoded by a nucleic acid that hybridizes to the Tkl1 nucleic acid (SEQ ID NO:66) or its complement under low stringency conditions,
(xxxiv) Tsa1 (SEQ ID NO:67), or a mammalian homolog thereof, or a variant of Tsa1 encoded by a nucleic acid that hybridizes to the Tsa1 nucleic acid (SEQ ID NO:68) or its complement under low stringency conditions,
(xxxv) Tye7 (SEQ ID NO:69), or a mammalian homolog thereof, or a variant of Tye7 encoded by a nucleic acid that hybridizes to the Tye7 nucleic acid (SEQ ID NO:70) or its complement under low stringency conditions,
(xxxvi) Vid24 (SEQ ID NO:71), or a mammalian homolog thereof, or a variant of Vid24 encoded by a nucleic acid that hybridizes to the Vid24 nucleic acid (SEQ ID NO:72) or its complement under low stringency conditions,
(xxxvii) Vps53 (SEQ ID NO:73), or a mammalian homolog thereof, or a variant of Vps53 encoded by a nucleic acid that hybridizes to the Vps53 nucleic acid (SEQ ID NO:74) or its complement under low stringency conditions,
(xxxviii) Ysh1 (SEQ ID NO:75), or a mammalian homolog thereof, or a variant of Ysh1 encoded by a nucleic acid that hybridizes to the Ysh1 nucleic acid (SEQ ID NO:76) or its complement under low stringency conditions,
(xxxix) Yth1 (SEQ ID NO:77), or a mammalian homolog thereof, or a variant of Yth1 encoded by a nucleic acid that hybridizes to the Yth1 nucleic acid (SEQ ID NO:78) or its complement under low stringency conditions,
(xl) Tcl046w (SEQ ID NO:79), or a mammalian homolog thereof, or a variant of Ycl046w encoded by a nucleic acid that hybridizes to the Tcl046w nucleic acid (SEQ ID NO:80) or its complement under low stringency conditions,
(xli) Ygr156w (SEQ ID NO:81), or a mammalian homolog thereof, or a variant of Ygr156w encoded by a nucleic acid that hybridizes to the Ygr156w nucleic acid (SEQ ID NO:82) or its complement under low stringency conditions,
(xlii) Yhl035c (SEQ ID NO:83), or a mammalian homolog thereof, or a variant of Yhl035c encoded by a nucleic acid that hybridizes to the Yhl035c nucleic acid (SEQ ID NO:84) or its complement under low stringency conditions,
(xliii) Ykl018w (SEQ ID NO:85), or a mammalian homolog thereof, or a variant of Ykl018w encoded by a nucleic acid that hybridizes to the Ykl018w nucleic acid (SEQ ID NO:86) or its complement under low stringency conditions,
(xliv) Ylr221c (SEQ ID NO:87), or a mammalian homolog thereof, or a variant of Ylr221c encoded by a nucleic acid that hybridizes to the Ylr221c nucleic acid (SEQ ID NO:88) or its complement under low stringency conditions,
(xlv) Ykl059c (SEQ ID NO:89), or a mammalian homolog thereof, or a variant of Ykl059c encoded by a nucleic acid that hybridizes to the Ykl059c nucleic acid (SEQ ID NO:90) or its complement under low stringency conditions,
(xlvi) Yml030w (SEQ ID NO:91), or a mammalian homolog thereof, or a variant of Yml030w encoded by a nucleic acid that hybridizes to the Yml030w nucleic acid (SEQ ID NO:92) or its complement under low stringency conditions, and
(xlvii) Yor179c (SEQ ID NO:93), or a mammalian homolog thereof, or a variant of Yor179c encoded by a nucleic acid that hybridizes to the Yor179c nucleic acid (SEQ ID NO:94) or its complement under low stringency conditions,
wherein said proteins are members of a native cellular Polyadenylation-complex, and
wherein said low stringency conditions comprise hybridization in a buffer comprising 35% formamide, 5×SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 ug/ml denatured salmon sperm DNA, and 10% (wt/vol) dextran sulfate for 18-20 hours at 40° C., washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 55° C., and washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 60° C.

46 more elements

A kit comprising in one or more containers

(a) an isolated first protein, or a functionally active fragment or functionally active derivative thereof, which first protein is selected from the group consisting of:
(i) Cft1 (SEQ ID NO:3), or a mammalian homolog thereof, or a variant of Cft1 encoded by a nucleic acid that hybridizes to the Cft1 nucleic acid (SEQ ID NO:4) or its complement under low stringency conditions,
(ii) Cft2 (SEQ ID NO:5), or a mammalian homolog thereof, or a variant of Cft2 encoded by a nucleic acid that hybridizes to the Cft2 nucleic acid (SEQ ID NO:6) or its complement under low stringency conditions,
(iii) Clp1 (SEQ ID NO:9), or a mammalian homolog thereof, or a variant of Clp1 encoded by a nucleic acid that hybridizes to the Clp1 nucleic acid (SEQ ID NO:10) or its complement under low stringency conditions,
(iv) Fip1 (SEQ ID NO:19), or a mammalian homolog thereof, or a variant of Fip1 encoded by a nucleic acid that hybridizes to the Fip1 nucleic acid (SEQ ID NO:20) or its complement under low stringency conditions,
(v) Pab1 (SEQ ID NO:33), or a mammalian homolog thereof, or a variant of Pab1 encoded by a nucleic acid that hybridizes to the Pab1 nucleic acid (SEQ ID NO:34) or its complement under low stringency conditions,
(vi) Pap1 (SEQ ID NO:35), or a mammalian homolog thereof, or a variant of Pap1 encoded by a nucleic acid that hybridizes to the Pap1 nucleic acid (SEQ ID NO:36) or its complement under low stringency conditions,
(vii) Pcf11 (SEQ ID NO:37), or a mammalian homolog thereof, or a variant of Pcf11 encoded by a nucleic acid that hybridizes to the Pcf11 nucleic acid (SEQ ID NO:38) or its complement under low stringency conditions,
(viii) Pfs2 (SEQ ID NO:43), or a mammalian homolog thereof, or a variant of Pfs2 encoded by a nucleic acid that hybridizes to the Pfs2 nucleic acid (SEQ ID NO:44) or its complement under low stringency conditions,
(ix) Pta1 (SEQ ID NO:45), or a mammalian homolog thereof, or a variant of Pta1 encoded by a nucleic acid that hybridizes to the Pta1 nucleic acid (SEQ ID NO:46) or its complement under low stringency conditions,
(x) Rna14 (SEQ ID NO:49), or a mammalian homolog thereof, or a variant of Rna14 encoded by a nucleic acid that hybridizes to the Rna14 nucleic acid (SEQ ID NO:50) or its complement under low stringency conditions,
(xi) Rna15 (SEQ ID NO:51), or a mammalian homolog thereof, or a variant of Rna15 encoded by a nucleic acid that hybridizes to the Rna15 nucleic acid (SEQ ID NO:52) or its complement under low stringency conditions,
(xii) Tif4632 (SEQ ID NO:63), or a mammalian homolog thereof, or a variant of Tif4632 encoded by a nucleic acid that hybridizes to the Tif4632 nucleic acid (SEQ ID NO:64) or its complement under low stringency conditions,
(xiii) Ykl059c (SEQ ID NO:89), or a mammalian homolog thereof, or a variant of Ykl059c encoded by a nucleic acid that hybridizes to the Ykl059c nucleic acid (SEQ ID NO:90) or its complement under low stringency conditions,
(xiv) Ysh1 (SEQ ID NO:75), or a mammalian homolog thereof, or a variant of Ysh1 encoded by a nucleic acid that hybridizes to the Ysh1 nucleic acid (SEQ ID NO:76) or its complement under low stringency conditions, and
(xv) Yth1 (SEQ ID NO:77), or a mammalian homolog thereof, or a variant of Yth1 encoded by a nucleic acid that hybridizes to the Yth1 nucleic acid (SEQ ID NO:78) or its complement under low stringency conditions;
and
(b) a second protein, or a functionally active fragment or functionally active derivative thereof, which second protein is selected from the group consisting of:
(i) Act1 (SEQ ID NO:1), or a mammalian homolog thereof, or a variant of Act1 encoded by a nucleic acid that hybridizes to the Act1 nucleic acid (SEQ ID NO:2) or its complement under low stringency conditions,
(ii) Cka1 (SEQ ID NO:7), or a mammalian homolog thereof, or a variant of Cka1 encoded by a nucleic acid that hybridizes to the Cka1 nucleic acid (SEQ ID NO:8) or its complement under low stringency conditions,
(iii) Eft2 (SEQ ID NO:11), or a mammalian homolog thereof, or a variant of Eft2 encoded by a nucleic acid that hybridizes to the Eft2 nucleic acid (SEQ ID NO:12) or its complement under low stringency conditions,
(iv) Eno2 (SEQ ID NO:13), or a mammalian homolog thereof, or a variant of Eno2 encoded by a nucleic acid that hybridizes to the Eno2 nucleic acid (SEQ ID NO:14) or its complement under low stringency conditions,
(v) Glc7 (SEQ ID NO:15), or a mammalian homolog thereof, or a variant of Glc7 encoded by a nucleic acid that hybridizes to the Glc7 nucleic acid (SEQ ID NO:16) or its complement under low stringency conditions,
(vi) Gpm1 (SEQ ID NO:17), or a mammalian homolog thereof, or a variant of Gpm1 encoded by a nucleic acid that hybridizes to the Gpm1 nucleic acid (SEQ ID NO:18) or its complement under low stringency conditions,
(vii) Hhf2 (SEQ ID NO:21), or a mammalian homolog thereof, or a variant of Hhf2 encoded by a nucleic acid that hybridizes to the Hhf2 nucleic acid (SEQ ID NO:22) or its complement under low stringency conditions,
(viii) Hta1 (SEQ ID NO:23), or a mammalian homolog thereof, or a variant of Hta1 encoded by a nucleic acid that hybridizes to the Hta1 nucleic acid (SEQ ID NO:24) or its complement under low stringency conditions,
(ix) Hsc82 (SEQ ID NO:25), or a mammalian homolog thereof, or a variant of Hsc82 encoded by a nucleic acid that hybridizes to the Hsc82 nucleic acid (SEQ ID NO:26) or its complement under low stringency conditions,
(x) Imd2 (SEQ ID NO:27), or a mammalian homolog thereof, or a variant of Imd2 encoded by a nucleic acid that hybridizes to the Imd2 nucleic acid (SEQ ID NO:28) or its complement under low stringency conditions,
(xi) Imd4 (SEQ ID NO:29), or a mammalian homolog thereof, or a variant of Imd4 encoded by a nucleic acid that hybridizes to the Imd4 nucleic acid (SEQ ID NO:30) or its complement under low stringency conditions,
(xii) Met6 (SEQ ID NO:31), or a mammalian homolog thereof, or a variant of Met6 encoded by a nucleic acid that hybridizes to the Met6 nucleic acid (SEQ ID NO:32) or its complement under low stringency conditions,
(xiii) Pdc1 (SEQ ID NO:39), or a mammalian homolog thereof, or a variant of Pdc1 encoded by a nucleic acid that hybridizes to the Pdc1 nucleic acid (SEQ ID NO:40) or its complement under low stringency conditions,
(xiv) Pfk1 (SEQ ID NO:41), or a mammalian homolog thereof, or a variant of Pfk1 encoded by a nucleic acid that hybridizes to the Pfk1 nucleic acid (SEQ ID NO:42) or its complement under low stringency conditions,
(xv) Ref2 (SEQ ID NO:47), or a mammalian homolog thereof, or a variant of Ref2 encoded by a nucleic acid that hybridizes to the Ref2 nucleic acid (SEQ ID NO:48) or its complement under low stringency conditions,
(xvi) Sec13 (SEQ ID NO:53), or a mammalian homolog thereof, or a variant of Sec13 encoded by a nucleic acid that hybridizes to the Sec13 nucleic acid (SEQ ID NO:54) or its complement under low stringency conditions,
(xvii) Sec31 (SEQ ID NO:55), or a mammalian homolog thereof, or a variant of Sec31 encoded by a nucleic acid that hybridizes to the Sec31 nucleic acid (SEQ ID NO:56) or its complement under low stringency conditions,
(xviii) Ssa3 (SEQ ID NO:57), or a mammalian homolog thereof, or a variant of Ssa3 encoded by a nucleic acid that hybridizes to the Ssa3 nucleic acid (SEQ ID NO:58) or its complement under low stringency conditions,
(xix) Ssu72 (SEQ ID NO:59), or a mammalian homolog thereof, or a variant of Ssu72 encoded by a nucleic acid that hybridizes to the Ssu72 nucleic acid (SEQ ID NO:60) or its complement under low stringency conditions,
(xx) Taf60 (SEQ ID NO:61), or a mammalian homolog thereof, or a variant of Taf60 encoded by a nucleic acid that hybridizes to the Taf60 nucleic acid (SEQ ID NO:62) or its complement under low stringency conditions,
(xxi) Tkl1 (SEQ ID NO:65), or a mammalian homolog thereof, or a variant of Tkl1 encoded by a nucleic acid that hybridizes to the Tkl1 nucleic acid (SEQ ID NO:66) or its complement under low stringency conditions,
(xxii) Tsa1 (SEQ ID NO:67), or a mammalian homolog thereof, or a variant of Tsa1 encoded by a nucleic acid that hybridizes to the Tsa1 nucleic acid (SEQ ID NO:68) or its complement under low stringency conditions,
(xxiii) Tye7 (SEQ ID NO:69), or a mammalian homolog thereof, or a variant of Tye7 encoded by a nucleic acid that hybridizes to the Tye7 nucleic acid (SEQ ID NO:70) or its complement under low stringency conditions,
(xxiv) Vid24 (SEQ ID NO:71), or a mammalian homolog thereof, or a variant of Vid24 encoded by a nucleic acid that hybridizes to the Vid24 nucleic acid (SEQ ID NO:72) or its complement under low stringency conditions,
(xxv) Vps53 (SEQ ID NO:73), or a mammalian homolog thereof, or a variant of Vps53 encoded by a nucleic acid that hybridizes to the Vps53 nucleic acid (SEQ ID NO:74) or its complement under low stringency conditions,
(xxvi) Ycl046w (SEQ ID NO:79), or a mammalian homolog thereof, or a variant of Ycl046w encoded by a nucleic acid that hybridizes to the Ycl046w nucleic acid (SEQ ID NO:80) or its complement under low stringency conditions,
(xxvii) Ygr156w (SEQ ID NO:81), or a mammalian homolog thereof, or a variant of Ygr156w encoded by a nucleic acid that hybridizes to the Ygr156w nucleic acid (SEQ ID NO:82) or its complement under low stringency conditions,
(xxviii) Yhl035c (SEQ ID NO:83), or a mammalian homolog thereof, or a variant of Yhl035c encoded by a nucleic acid that hybridizes to the Yhl035c nucleic acid (SEQ ID NO:84) or its complement under low stringency conditions,
(xxix) Ykl018w (SEQ ID NO:85), or a mammalian homolog thereof, or a variant of Ykl018w encoded by a nucleic acid that hybridizes to the Ykl018w nucleic acid (SEQ ID NO:86) or its complement under low stringency conditions,
(xxx) Ylr221c (SEQ ID NO:87), or a mammalian homolog thereof, or a variant of Ylr221c encoded by a nucleic acid that hybridizes to the Ylr221c nucleic acid (SEQ ID NO:88) or its complement under low stringency conditions,
(xxxi) Yml030w (SEQ ID NO:91), or a mammalian homolog thereof, or a variant of Yml030w encoded by a nucleic acid that hybridizes to the Yml030w nucleic acid (SEQ ID NO:92) or its complement under low stringency conditions, and
(xxxii) Yor179c (SEQ ID NO:93), or a mammalian homolog thereof, or a variant of Yor179c encoded by a nucleic acid that hybridizes to the Yor179c nucleic acid (SEQ ID NO:94) or its complement under low stringency conditions,
wherein said first protein and said second protein are members of a native cellular Polyadenylation-complex, and wherein said low stringency conditions comprise hybridization in a buffer comprising 35% formamide, 5×SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 ug/ml denatured salmon sperm DNA, and 10% (wt/vol) dextran sulfate for 18-20 hours at 40° C., washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 55° C., and washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 60° C.

48 more elements

Component of the Polyadenylation-complex selected from

a) yeast proteins
(i) Tcl046w (SEQ ID NO:59),
(ii) Ygr156w (SEQ ID NO:61),
(iii) Yhl035c (SEQ ID NO:63),
(iv) Ykl018w (SEQ ID NO:179),
(v) Ylr221c (SEQ ID NO:67),
(vi) Yml030w (SEQ ID NO:69), and
(vii) Yor179c (SEQ ID NO:71).
b) the mammalian homologs/orthologs of the proteins of (a), and
c) a functionally active fragment or functionally active derivate of the proteins according to (a) and (b) carrying one or more amino acid substitutions, deletions and/or additions.

7 more elements

Dependencies are inferred from the claim text.

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Abstract

An isolated complex selected from complex (I) and comprising

(a) a first protein, or a functionally active fragment or functionally active derivative thereof, which first protein is selected from the group consisting of:
(i) Cft1 (SEQ ID NO:3), or a mammalian homolog thereof, or a variant of Cft1 encoded by a nucleic acid that hybridizes to the Cft1 nucleic acid (SEQ ID NO:4) or its complement under low stringency conditions,
(ii) Cft2 (SEQ ID NO:5), or a mammalian homolog thereof, or a variant of Cft2 encoded by a nucleic acid that hybridizes to the Cft2 nucleic acid (SEQ ID NO:6) or its complement under low stringency conditions,
(iii) Clp1 (SEQ ID NO:9), or a mammalian homolog thereof, or a variant of Clp1 encoded by a nucleic acid that hybridizes to the Clp1 nucleic acid (SEQ ID NO:10) or its complement under low stringency conditions,
(iv) Fip1 (SEQ ID NO:19), or a mammalian homolog thereof, or a variant of Fip1 encoded by a nucleic acid that hybridizes to the Fip1 nucleic acid (SEQ ID NO:20) or its complement under low stringency conditions,
(v) Pab1 (SEQ ID NO:33), or a mammalian homolog thereof, or a variant of Pab1 encoded by a nucleic acid that hybridizes to the Pab1 nucleic acid (SEQ ID NO:34) or its complement under low stringency conditions,
(vi) Pap1 (SEQ ID NO:35), or a mammalian homolog thereof, or a variant of Pap1 encoded by a nucleic acid that hybridizes to the Pap1 nucleic acid (SEQ ID NO:36) or its complement under low stringency conditions,
(vii) Pcf11 (SEQ ID NO:37), or a mammalian homolog thereof, or a variant of Pcf11 encoded by a nucleic acid that hybridizes to the Pcf11 nucleic acid (SEQ ID NO:38) or its complement under low stringency conditions,
(viii) Pfs2 (SEQ ID NO:43), or a mammalian homolog thereof, or a variant of Pfs2 encoded by a nucleic acid that hybridizes to the Pfs2 nucleic acid (SEQ ID NO:44) or its complement under low stringency conditions,
(ix) Pta1 (SEQ ID NO:45), or a mammalian homolog thereof, or a variant of Pta1 encoded by a nucleic acid that hybridizes to the Pta1 nucleic acid (SEQ ID NO:46) or its complement under low stringency conditions,
(x) Rna14 (SEQ ID NO:49), or a mammalian homolog thereof, or a variant of Rna14 encoded by a nucleic acid that hybridizes to the Rna14 nucleic acid (SEQ ID NO:50) or its complement under low stringency conditions,
(xi) Rna15 (SEQ ID NO:51), or a mammalian homolog thereof, or a variant of Rna15 encoded by a nucleic acid that hybridizes to the Rna15 nucleic acid (SEQ ID NO:52) or its complement under low stringency conditions,
(xii) Tif4632 (SEQ ID NO:63), or a mammalian homolog thereof, or a variant of Tif4632 encoded by a nucleic acid that hybridizes to the Tif4632 nucleic acid (SEQ ID NO:64) or its complement under low stringency conditions,
(xiii) Ykl059c (SEQ ID NO:89), or a mammalian homolog thereof, or a variant of Ykl059c encoded by a nucleic acid that hybridizes to the Ykl059c nucleic acid (SEQ ID NO:90) or its complement under low stringency conditions,
(xiv) Ysh1 (SEQ ID NO:75), or a mammalian homolog thereof, or a variant of Ysh1 encoded by a nucleic acid that hybridizes to the Ysh1 nucleic acid (SEQ ID NO:76) or its complement under low stringency conditions, and
(xv) Yth1 (SEQ ID NO:77), or a mammalian homolog thereof, or a variant of Yth1 encoded by a nucleic acid that hybridizes to the Yth1 nucleic acid (SEQ ID NO:78) or its complement under low stringency conditions;
and
(b) a second protein, or a functionally active fragment or functionally active derivative thereof, which second protein is selected from the group consisting of:
(i) Act1 (SEQ ID NO:1), or a mammalian homolog thereof, or a variant of Act1 encoded by a nucleic acid that hybridizes to the Act1 nucleic acid (SEQ ID NO:2) or its complement under low stringency conditions,
(ii) Cka1 (SEQ ID NO:7), or a mammalian homolog thereof, or a variant of Cka1 encoded by a nucleic acid that hybridizes to the Cka1 nucleic acid (SEQ ID NO:8) or its complement under low stringency conditions,
(iii) Eft2 (SEQ ID NO:11), or a mammalian homolog thereof, or a variant of Eft2 encoded by a nucleic acid that hybridizes to the Eft2 nucleic acid (SEQ ID NO:12) or its complement under low stringency conditions,
(iv) Eno2 (SEQ ID NO:13), or a mammalian homolog thereof, or a variant of Eno2 encoded by a nucleic acid that hybridizes to the Eno2 nucleic acid (SEQ ID NO:14) or its complement under low stringency conditions,
(v) Glc7 (SEQ ID NO:15), or a mammalian homolog thereof, or a variant of Glc7 encoded by a nucleic acid that hybridizes to the Glc7 nucleic acid (SEQ ID NO:16) or its complement under low stringency conditions,
(vi) Gpm1 (SEQ ID NO:17), or a mammalian homolog thereof, or a variant of Gpm1 encoded by a nucleic acid that hybridizes to the Gpm1 nucleic acid (SEQ ID NO:18) or its complement under low stringency conditions,
(vii) Hhf2 (SEQ ID NO:21), or a mammalian homolog thereof, or a variant of Hhf2 encoded by a nucleic acid that hybridizes to the Hhf2 nucleic acid (SEQ ID NO:22) or its complement under low stringency conditions,
(viii) Hta1 (SEQ ID NO:23), or a mammalian homolog thereof, or a variant of Hta1 encoded by a nucleic acid that hybridizes to the Hta1 nucleic acid (SEQ ID NO:24) or its complement under low stringency conditions,
(ix) Hsc82 (SEQ ID NO:25), or a mammalian homolog thereof, or a variant of Hsc82 encoded by a nucleic acid that hybridizes to the Hsc82 nucleic acid (SEQ ID NO:26) or its complement under low stringency conditions,
(x) Imd2 (SEQ ID NO:27), or a mammalian homolog thereof, or a variant of Imd2 encoded by a nucleic acid that hybridizes to the Imd2 nucleic acid (SEQ ID NO:28) or its complement under low stringency conditions,
(xi) Imd4 (SEQ ID NO:29), or a mammalian homolog thereof, or a variant of Imd4 encoded by a nucleic acid that hybridizes to the Imd4 nucleic acid (SEQ ID NO:30) or its complement under low stringency conditions,
(xii) Met6 (SEQ ID NO:31), or a mammalian homolog thereof, or a variant of Met6 encoded by a nucleic acid that hybridizes to the Met6 nucleic acid (SEQ ID NO:32) or its complement under low stringency conditions,
(xiii) Pdc1 (SEQ ID NO:39), or a mammalian homolog thereof, or a variant of Pdc1 encoded by a nucleic acid that hybridizes to the Pdc1 nucleic acid (SEQ ID NO:40) or its complement under low stringency conditions,
(xiv) Pfk1 (SEQ ID NO:41), or a mammalian homolog thereof, or a variant of Pfk1 encoded by a nucleic acid that hybridizes to the Pfk1 nucleic acid (SEQ ID NO:42) or its complement under low stringency conditions,
(xv) Ref2 (SEQ ID NO:47), or a mammalian homolog thereof, or a variant of Ref2 encoded by a nucleic acid that hybridizes to the Ref2 nucleic acid (SEQ ID NO:48) or its complement under low stringency conditions,
(xvi) Sec13 (SEQ ID NO:53), or a mammalian homolog thereof, or a variant of Sec13 encoded by a nucleic acid that hybridizes to the Sec13 nucleic acid (SEQ ID NO:54) or its complement under low stringency conditions,
(xvii) Sec31 (SEQ ID NO:55), or a mammalian homolog thereof, or a variant of Sec31 encoded by a nucleic acid that hybridizes to the Sec31 nucleic acid (SEQ ID NO:56) or its complement under low stringency conditions,
(xviii) Ssa3 (SEQ ID NO:57), or a mammalian homolog thereof, or a variant of Ssa3 encoded by a nucleic acid that hybridizes to the Ssa3 nucleic acid (SEQ ID NO:58) or its complement under low stringency conditions,
(xix) Ssu72 (SEQ ID NO:59), or a mammalian homolog thereof, or a variant of Ssu72 encoded by a nucleic acid that hybridizes to the Ssu72 nucleic acid (SEQ ID NO:60) or its complement under low stringency conditions,
(xx) Taf60 (SEQ ID NO:61), or a mammalian homolog thereof, or a variant of Taf60 encoded by a nucleic acid that hybridizes to the Taf60 nucleic acid (SEQ ID NO:62) or its complement under low stringency conditions,
(xxi) Tkl1 (SEQ ID NO:65), or a mammalian homolog thereof, or a variant of Tkl1 encoded by a nucleic acid that hybridizes to the Tkl1 nucleic acid (SEQ ID NO:66) or its complement under low stringency conditions,
(xxii) Tsa1 (SEQ ID NO:67), or a mammalian homolog thereof, or a variant of Tsa1 encoded by a nucleic acid that hybridizes to the Tsa1 nucleic acid (SEQ ID NO:68) or its complement under low stringency conditions,
(xxiii) Tye7 (SEQ ID NO:69), or a mammalian homolog thereof, or a variant of Tye7 encoded by a nucleic acid that hybridizes to the Tye7 nucleic acid (SEQ ID NO:70) or its complement under low stringency conditions,
(xxiv) Vid24 (SEQ ID NO:71), or a mammalian homolog thereof, or a variant of Vid24 encoded by a nucleic acid that hybridizes to the Vid24 nucleic acid (SEQ ID NO:72) or its complement under low stringency conditions,
(xxv) Vps53 (SEQ ID NO:73), or a mammalian homolog thereof, or a variant of Vps53 encoded by a nucleic acid that hybridizes to the Vps53 nucleic acid (SEQ ID NO:74) or its complement under low stringency conditions,
(xxvi) Ycl046w (SEQ ID NO:79), or a mammalian homolog thereof, or a variant of Ycl046w encoded by a nucleic acid that hybridizes to the Ycl046w nucleic acid (SEQ ID NO:80) or its complement under low stringency conditions,
(xxvii) Ygr156w (SEQ ID NO:81), or a mammalian homolog thereof, or a variant of Ygr156w encoded by a nucleic acid that hybridizes to the Ygr156w nucleic acid (SEQ ID NO:82) or its complement under low stringency conditions,
(xxviii) Yhl035c (SEQ ID NO:83), or a mammalian homolog thereof, or a variant of Yhl035c encoded by a nucleic acid that hybridizes to the Yhl035c nucleic acid (SEQ ID NO:84) or-its complement under low stringency conditions,
(xxix) Ykl018w (SEQ ID NO:85), or a mammalian homolog thereof, or a variant of Ykl018w encoded by a nucleic acid that hybridizes to the Ykl018w nucleic acid (SEQ ID NO:86) or its complement under low stringency conditions,
(xxx) Ylr221c (SEQ ID NO:87), or a mammalian homolog thereof, or a variant of Ylr221c encoded by a nucleic acid that hybridizes to the Ylr221c nucleic acid (SEQ ID NO:88) or its complement under low stringency conditions,
(xxxi) Yml030w (SEQ ID NO:91), or a mammalian homolog thereof, or a variant of Yml030w encoded by a nucleic acid that hybridizes to the Yml030w nucleic acid (SEQ ID NO:92) or its complement under low stringency conditions, and
(xxxii) Yor179c (SEQ ID NO:93), or a mammalian homolog thereof, or a variant of Yor179c encoded by a nucleic acid that hybridizes to the Yor179c nucleic acid (SEQ ID NO:94) or its complement under low stringency conditions,
wherein said first protein and said second protein are members of a native cellular Polyadenylation-complex, and wherein said low stringency conditions comprise hybridization in a buffer comprising 35% formamide, 5×SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 ug/ml denatured salmon sperm DNA, and 10% (wt/vol) dextran sulfate for 18-20 hours at 40° C., washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 55° C., and washing in a buffer consisting of 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS for 1.5 hours at 60° C., and a complex (II) comprising at least two second proteins.

48 more elements