Kinesin-like proteins and methods of use
The invention relates to methods and compounds for treating subjects who have proliferative disorders. The invention also relates to screening methods for identifying such compounds. The methods and compositions target the activity of the human kinesin-like protein KIF18A (and the KIF18A ortholog, KLP67A) and homologs thereof, and a gene encoding KIF18A or a homologous or orthologous gene. KIF18A and KLP67A are kinesin superfamily (KIF) proteins. Members of the KIF protein family are known to participate in chromosomal and spindle movements during mitosis and meiosis thereby making them attractive targets for treating cancers and the like.
1. A method for identifying a compound that modulates activity of a KIF18A polypeptide, the method comprising:
a) obtaining a test cell containing a KIF18A polypeptide that is at least 95% identical to SEQ ID NO:2 and that localizes to the distal ends of astral microtubules in dividing cells, and a control cell containing said KIF18A polypeptide;
b) incubating the test cell with a test compound; and
c) detecting an altered localization of the KIF18A polypeptide in the test cell as compared to the KIF18A polypeptide in the control cell, wherein the altered localization is localization to a region other than a growth cone of an interphase cell or the distal ends of astral microtubules of a dividing cell,
wherein an altered localization indicates that the test compound modulates activity of the KIF18A polypeptide.
2. The method of claim 1 , wherein the KIF18A polypeptide has comprises the sequence of SEQ ID NO:2.
3. The method of claim 1 , wherein the test compound is an antisense nucleic acid molecule, a small inhibitory RNA (siRNA), a ribozyme, a triple helix molecule, an antibody, a polypeptide, a peptoid, a polypeptide mimetic, a small inorganic molecule, or a small non-nucleic acid organic molecule.
4. The method of claim 1 , wherein the polypeptide is localized to a region of a dividing cell other than the distal ends of astral microtubules in the presence of the test compound.
5. The method of claim 1 , wherein the polypeptide is localized using immunocytochemistry.
6. The method of claim 1 , wherein the polypeptide is fused to a reporter molecule.
7. The method of claim 6 , wherein the reporter molecule is green fluorescent protein (GFP), β-glucuronidase (GUS), luciferase, chloramphenicol transacetylase (CAT), horseradish peroxidase (HRP), or β-galactosidase.
8. The method of claim 2 , wherein the KIF18A polypeptide consists of the sequence of SEQ ID NO:2.
9. The method of claim 1 , further comprising assaying spindle assembly or function of the test cell as compared to the control cell.
10. The method of claim 1 , further comprising assaying cell division in the test cell as compared to the control cell.
11. The method of claim 1 , further comprising determining whether the test compound binds to the KIF18A polypeptide.
12. The method of claim 7 , wherein the reporter molecule is green fluorescent protein (GFP).
13. The method of claim 7 , wherein the reporter molecule is β-glucuronidase (GUS).
14. The method of claim 7 , wherein the reporter molecule is luciferase.
15. The method of claim 7 , wherein the reporter molecule is chloramphenicol transacetylase (CAT).
16. The method of claim 7 , wherein the reporter molecule is horseradish peroxidase (HRP).
17. The method of claim 7 , wherein the reporter molecule is β-galactosidase.