Novel RGS9 protein binding interactions and methods of use thereof

A method for assaying the effects of test compounds on the activity of a RGS9-evectin polypeptide dimer comprising the steps of:

• (a) providing recombinant cells comprising a RGS9 polypeptide having an amino acid sequence of SEQ ID NO:2 and an evectin polypeptide having an amino acid sequence of SEQ ID NO:4;
• (b) contacting the cells with a test compound;
• and
• (c) determining the effect of the test compound on the activity of the RGS9-evectin dimer in the presence and absence of the test compound.

A method for assaying the effects of test compounds on the activity of a RGS9-evectin polypeptide dimer comprising the steps of:

• (a) providing a transgenic animal comprising a polynucleotide encoding a RGS9 polypeptide having an amino acid sequence of SEQ ID NO:2 and a polynucleotide encoding an evectin polypeptide having an amino acid sequence of SEQ ID NO:4;
• (b) administering a test compound to the animal;
• and
• (c) determining the effects of the test compound on the activity of the RGS9-evectin polypeptide in the presence and absence of the test compound.

A method for assaying the effects of test compounds on a transgenic animal with a genome comprising a functional disruption of the evectin binding domain in the RGS9 polypeptide, the method comprising:

• (a) providing a transgenic animal whose genome comprises a disruption of the endogenous polynucleotide encoding the RGS9 polypeptide, wherein the disruption occurs in the evectin binding domain;
• (b) administering a test compound to the animal;
• and
• (c) determining the effects of the test compound on the activity of the RGS9 polypeptide in the presence and absence of the test compound.

A method for assaying the effects of test compounds on a transgenic animal with a genome comprising a functional disruption of the RGS9 binding domain in the evectin polypeptide, the method comprising:

• (b) providing a transgenic animal whose genome comprises a disruption of the endogenous polynucleotide encoding the evectin polypeptide, wherein the disruption occurs in the RGS9 binding domain;
• (b) administering a test compound to the animal;
• and
• (c) determining the effects of the test compound on the activity of the evectin polypeptide in the presence and absence of the test compound.

A method for assaying the effects of test compounds on the binding interaction of RGS9 and evectin polypeptides comprising the steps of:

• (a) providing yeast cells for a yeast two-hybrid system comprising a RGS9 polypeptide having an amino acid sequence of SEQ ID NO:2 and an evectin polypeptide having an amino acid sequence of SEQ ID NO:4;
• (b) contacting the cells with a test compound;
• and
• (c) determining the effect of the test compound on the binding interaction of the RGS9 and evectin polypeptides in the presence and absence of the test compound.

A method for the treatment of a subject in need of inhibiting RGS9-evectin dimer activity comprising:

• (a) administering to the subject a therapeutically effective amount of an antagonist to the RGS9-evectin;
• or
• (b) administering to the subject a polynucleotide that inhibits the expression of a polynucleotide encoding a RGS9-evectin polypeptide;
• or
• (c) administering to the subject a therapeutically effective amount of a polypeptide that competes with RGS9-evectin for its ligand.

A method for producing a transgenic animal whose genome comprises a functional disruption of the evectin binding domain in a polynucleotide encoding a RGS9 polypeptide, the method comprising:

• (a) providing a polynucleotide encoding a RGS9 polypeptide having a functional disruption in the evectin binding domain, wherein the binding domain comprises the amino acid sequence of amino acid 461 through amino acid 602 of SEQ ID NO:2;
• (b) introducing the disrupted polynucleotide into embryonic stem cells;
• (c) selecting those embryonic stem cells that comprise the disrupted polynucleotide;
• (d) introducing an embryonic stem cell of step (c) into a blastocyst;
• transferring the blastocyst of step (d) to a pseudopregnant animal;
• and
• (e) allowing the transferred blastocyst to develop into an animal chimeric for the disruption.

A method for producing a transgenic animal whose genome comprises a functional disruption of the RGS9 binding domain in a polynucleotide encoding an evectin polypeptide, the method comprising:

• (a) providing a polynucleotide encoding an evectin polypeptide having a functional disruption in the RGS9 binding domain, wherein the binding domain comprises the amino acid sequence of amino acid 79 through amino acid 136 of SEQ ID NO:4;
• (b) introducing the disrupted polynucleotide into embryonic stem cells;
• (c) selecting those embryonic stem cells that comprise the disrupted polynucleotide;
• (d) introducing an embryonic stem cell of step (c) into a blastocyst;
• transferring the blastocyst of step (d) to a pseudopregnant animal;
• and
• (e) allowing the transferred blastocyst to develop into an animal chimeric for the disruption.