DNA fragmentation factor involved in apoptosis
AssigneeBoard of Regents, The University of Texas System
InventorsXiaodong Wang, Xueson Liu
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Abstract
The invention provides methods and compositions relating to DNA Fragmentation Factor (DFF) polypeptides and related nucleic acids. More particularly, the present invention provides the sequence for the active subunit of DFF. The polypeptides may be produced recombinantly from host cells transformed from the disclosed DFF encoding nucleic acids or purified from human cells. The invention provides isolated DFF hybridization probes and primers capable of specifically hybridization with the disclosed DFF genes, DFF-specific binding agents such as specific antibodies, and methods of making and using the subject compositions.
Claims100
An isolated polypeptide encoding a DFF40 DNA fragmentation factor.
An isolated peptide having between about 10 and about 50 consecutive residues of a DFF40 DNA fragmentation factor.
A monoclonal antibody that binds immunologically to a DFF40 DNA fragmentation factor.
A hybridoma cell that produces a monoclonal antibody that binds immunologically to a DFF40 DNA fragmentation factor.
A polyclonal antisera, antibodies of which bind immunologically to a DFF40 DNA fragmentation factor.
An isolated nucleic acid comprising a region, or the complement thereof, encoding a DFF40 DNA fragmentation factor or an allelic variant thereof.
An isolated oligonucleotide of between about 15 and about 50 consecutive bases of a nucleic acid, or complement thereof, encoding a DFF40 DNA fragmentation factor.
A plasmid construct comprising a first nucleic acid encoding a DFF40 DNA fragmentation factor.
A method of inducing apoptosis in a cell comprising the step of providing said cell with a DFF40 DNA fragmentation factor, wherein the provision of said DFF40 to said cell results in apoptosis.
A method for inhibiting the growth of a cancer cell comprising the step of contacting a cancer cell with a DNA fragmentation factor designated DFF40 under conditions permitting the uptake of said DNA fragmentation factor by said cell, wherein the presence of said DFF40 in said cell induces apoptosis.
A method for treating cancer comprising the step of contacting a tumor cell within a subject with a nucleic acid (i) encoding a DFF40 DNA fragmentation factor and (ii) a promoter active in said tumor cell, wherein said promoter is operably linked to the region encoding said DNA fragmentation factor, under conditions permitting the uptake of said nucleic acid by said tumor cell.
A method of identifying a modulator of DFF40 activity comprising the steps of:
- (i) providing a cell expressing a DFF40/DFF45 complex;
- (ii) contacting said cell with a candidate substance;
- (iii) activating DFF40;
- and
- (iv) comparing the apoptosis of the cell in step (iii) with the apoptosis observed when said candidate substance is not added,
- wherein an alteration in apoptosis indicates that said candidate substance is a modulator said apoptotic activity.
A modulator of apoptotic activity identified according to a method comprising the steps of:
- (i) providing a cell expressing a DFF40/DFF45 complex;
- (ii) contacting said cell with a candidate substance;
- (iii) activating DFF40;
- and
- (iv) comparing the apoptosis of the cell in step (iii) with the apoptosis observed when said candidate substance is not added,
- wherein an alteration in apoptosis indicates that said candidate substance is a modulator said apoptotic activity.
An isolated DNA fragmentation factor complex for regulating chromatin stability, said complex comprising a DFF40 polypeptide and a DFF45 polypeptide.
A method of producing a functional DNA fragmentation factor comprising:
- (i) providing to a cell
- (a) a first nucleic acid encoding a DFF40 polypeptide;
- and
- (b) a second nucleic acid encoding a DFF45 polypeptide;
- and
- (ii) expressing said complex in a cell,
- wherein the coexpression of said polypeptides allows for the formation of a functional DFF40 polypeptide.
Dependencies are inferred from the claim text.