Essential fungal genes and their use
AssigneeMillennium Pharmaceuticals, Inc., a Massachusetts corporation
InventorsYigal Koltin, Victoria Gavrias
Life of this patent
Estimating the term…
Abstract
Disclosed are essential Aspergillus polypeptides and genes (AN97, AN17, AN80, and AN85), as well as homologs thereof, which can be used to identify antifungal agents for treating fungal infections such as aspergillosis.
Claims66
An isolated nucleic acid or an allelic variant thereof encoding:
- an AN97 polypeptide comprising the amino acid sequence of SEQ ID NO:1, as depicted in FIG. 1;
- an AN80 polypeptide comprising the amino acid sequence of SEQ ID NO:3, as depicted in FIG. 2;
- an AN85 polypeptide comprising the amino acid sequence of SEQ ID NO:5, as depicted in FIG. 3;
- or an AN17 polypeptide comprising the amino acid sequence of SEQ ID NO:7, as depicted in FIG. 4.
An isolated nucleic acid comprising a sequence selected from the group consisting of:
- (a) SEQ ID NO:2, as depicted in FIG. 1, or degenerate variants thereof;
- (b) SEQ ID NO:2, or degenerate variants thereof, wherein T is replaced by U;
- (c) nucleic acids complementary to (a) and (b);
- (d) fragments of (a), (b), and (c) that are at least 15 base pairs in length and which hybridize under stringent conditions to genomic DNA encoding the polypeptide of SEQ ID NO:1;
- (e) SEQ ID NO:4, as depicted in FIG. 2, or degenerate variants thereof;
- (f) SEQ ID NO:4, or degenerate variants thereof, wherein T is replaced by U;
- (g) nucleic acids complementary to (e) and (f);
- (h) fragments of (e), (f), and (g) that are at least 15 base pairs in length and which hybridize under stringent conditions to genomic DNA encoding the polypeptide of SEQ ID NO:3;
- (i) SEQ ID NO:6, as depicted in FIG. 3, or degenerate variants thereof;
- (j) SEQ ID NO:6, or degenerate variants thereof, wherein T is replaced by U;
- (k) nucleic acids complementary to (i) and (j);
- (1) fragments of (i), (j), and (k) that are at least 15 base pairs in length and which hybridize under stringent conditions to genomic DNA encoding the polypeptide of SEQ ID NO:5
- (m) SEQ ID NO:8, as depicted in FIG. 4, or degenerate variants thereof;
- (n) SEQ ID NO:8, or degenerate variants thereof, wherein T is replaced by U;
- (o) nucleic acids complementary to (m) and (n);
- and
- (p) fragments of (m), (n), and (o) that are at least 15 base pairs in length and which hybridize under stringent conditions to genomic DNA encoding the polypeptide of SEQ ID NO:7.
An isolated nucleic acid from Aspergillus comprising a nucleotide sequence that is at least 85% identical to a nucleotide sequence selected from the group consisting of
- SEQ ID NO:2, and encoding an AN97 polypeptide;
- SEQ ID NO:4, and encoding an AN80 polypeptide;
- SEQ ID NO:6, and encoding an AN85 polypeptide;
- and
- SEQ ID NO:8, and encoding an AN17 polypeptide.
An isolated nucleic acid that is at least 15 base pairs in length and hybridizes under stringent conditions to a nucleotide sequence selected from the group consisting of
- SEQ ID NO:2;
- SEQ ID NO:4;
- SEQ ID NO:6;
- and
- SEQ ID NO:8.
An isolated nucleic acid molecule, said molecule comprising the cDNA sequence contained within an American Type Culture Collection (ATCC) accession number selected from the group consisting of ______, ______, ______, and ______.
A polypeptide comprising an amino acid sequence selected from the group consisting of:
- the amino acid sequence of SEQ ID NO:1, as depicted in FIG. 1;
- the amino acid sequence of SEQ ID NO:3, as depicted in FIG. 2;
- the amino acid sequence of SEQ ID NO:5, as depicted in FIG. 3;
- and
- the amino acid sequence of SEQ ID NO:7, as depicted in FIG. 4.
A method for identifying an antifungal agent, the method comprising:
- (a) contacting an AN polypeptide with a test compound, wherein the AN polypeptide is selected from the group consisting of AN97, AN17, AN85, and AN80;
- (b) detecting binding of the test compound to the AN polypeptide;
- and
- (c) determining whether a test compound that binds to an AN polypeptide inhibits growth of fungi, relative to growth of fungi cultured in the absence of a test compound that binds to an AN polypeptide, wherein inhibition of growth is an indication that the test compound is an antifungal agent.
A pharmaceutical formulation comprising an antifungal agent, wherein the agent is a ribozyme that inhibits the function of AN97, AN17, AN80, or AN85.
A pharmaceutical formulation comprising an antifungal agent, wherein the agent is an antisense nucleic acid that inhibits the function of AN97, AN17, AN80, or AN85.
A method for identifying an antifungal agent, the method comprising:
- (a) contacting an AN polypeptide with a test compound, wherein the AN polypeptide is selected from the group consisting of AN97, AN17, AN85, and AN80;
- (b) detecting a decrease in function of the AN polypeptide contacted with the test compound;
- and
- (c) determining whether a test compound that decreases function of a contacted AN polypeptide inhibits growth of fungi, relative to growth of fungi cultured in the absence of a test compound that decreases function of a contacted AN polypeptide, wherein inhibition of growth is an indication that the test compound is an antifungal agent.
A method for identifying an antifungal agent, the method comprising:
- (a) contacting a nucleic acid encoding an AN polypeptide with a test compound, wherein the AN polypeptide is selected from the group consisting of AN97, AN17, AN80, and AN85;
- (b) detecting binding of the test compound to the nucleic acid;
- and
- (c) determining whether a test compound that binds the nucleic acid inhibits growth of fungi, relative to growth of fungi cultured in the absence of the test compound that binds the nucleic acid, wherein inhibition of growth is an indication that the test compound is an antifungal agent.
A method for identifying an anti-yeast agent, the method comprising:
- (a) contacting a yeast homolog of an AN polypeptide with a test compound, wherein the AN polypeptide is selected from the group consisting of AN97, AN17, AN85, and AN80;
- (b) detecting binding of the test compound to the yeast homolog;
- and
- (c) determining whether a test compound that binds to the yeast homolog inhibits growth of yeast, relative to growth of yeast cultured in the absence of the test compound that binds the yeast homolog, wherein inhibition of growth is indication that the test compound is an anti-yeast agent.
A method for identifying an anti-yeast agent, the method comprising:
- (a) contacting a homolog of an AN polypeptide with a test compound, wherein the AN polypeptide is selected from the group consisting of AN97, AN17, AN85, and AN80;
- (b) detecting a decrease in function of the AN polypeptide contacted by the homolog;
- and
- (c) determining whether a test compound that decreases function of a contacted AN polypeptide inhibits growth of yeast, relative to growth of yeast cultured in the absence of a test compound that decreases function of a contacted AN polypeptide, wherein inhibition of growth is an indication that the test compound is an anti-yeast agent.
A method for identifying an anti-yeast agent, the method comprising:
- (a) contacting a nucleic acid encoding a homolog of an AN polypeptide with a test compound, wherein the AN polypeptide is selected from the group consisting of AN97, AN17, AN80, and AN85;
- (b) detecting binding of the test compound to the nucleic acid;
- and
- (c) determining whether a test compound that binds the nucleic acid inhibits growth of yeast, relative to growth of yeast cultured in the absence of a test compound that binds the nucleic acid, wherein inhibition of growth is an indication that the test compound is an anti-yeast agent.
Dependencies are inferred from the claim text.