Compositions, organisms and methodologies employing a novel human kinase

• US 20050059025A1
• Filed: 10/07/2003
• Published: 03/17/2005
• Est. Priority Date: 10/07/2002
• Status: Abandoned Application

• Current Assignee: Liu Wei, Wyeth (Pfizer Inc.)
• Original Assignee: Five Giralda Farms
• Inventors: Liu, Wei, Wu, Leeying
• Application Number:
  US10/679,366
• Publication Number:
  US 20050059025A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12N 9/1205 Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases

Abstract

This invention provides compositions, organisms and methodologies employing a novel human protein kinase, MCRK2. The novel human kinase has sequence homology to rat myotonic dystrophy kinase-related Cdc42 binding kinase (MRCK) alpha. The gene encoding the novel kinase is localized in locus 1q42 of human chromosome 1. The novel protein kinase comprises multiple functional/structural domains that include a kinase domain, a pkinase_C domain, a DAG-PE binding domain, and a CNH domain. The sequence and structure similarity between the novel human protein and rat MRCK alpha indicates that the novel human protein may function as a downstream effector of Cdc42 in cytoskeleton reorganization.

Claims

1. An isolated polynucleotide comprising a nucleic acid sequence which encodes the amino acid sequence depicted in SEQ ID NO:

2.

2. The polynucleotide according to claim 1, wherein the nucleic acid sequence is selected from the group consisting of:

(a) the nucleic acid sequence as shown in SEQ ID NO;

1;

(b) the complement of (a); and

(c) a nucleic acid sequence that differs from (a) or (b) due to the degeneracy of the genetic code.

3. The polynucleotide according to claim 1, wherein the nucleic acid sequence is selected from the group consisting of:

(a) the nucleic acid sequence as shown in SEQ ID NO;

3;

(b) the complement of (a); and

(c) a nucleic acid sequence that differs from (a) or (b) due to the degeneracy of the genetic code.

14. A host cell containing the polynucleotide of claim 1 or a variant thereof.

15. A transgenic non-human animal comprising the polynucleotide of claim 1 or a variant thereof.

4. An isolated polynucleotide comprising a variant of a nucleic acid sequence, wherein said nucleic acid sequence encodes the amino acid sequence depicted in SEQ ID NO:

2, and wherein the variant and said nucleic acid sequence have at least 96% sequence identity.

5. The polynucleotide according to claim 4, wherein the variant and said nucleic acid sequence have at least 98% sequence identity.

6. An isolated polynucleotide that is capable of hybridizing under highly stringent conditions to the sequence set forth in SEQ ID NO:

1, or the complement thereof, wherein said polynucleotide encodes a protein having MRCK activity.

7. The polynucleotide of claim 6, wherein said polynucleotide and the sequence set forth in SEQ ID NO:

1 have a sequence identity of at least 96%.

8. An isolated polypeptide comprising a fragment of SEQ ID NO:

2, wherein said fragment comprises at least 500 consecutive amino acid residues of SEQ ID NO;

2.

9. The polypeptide according to claim 8, wherein the fragment consists of SEQ ID NO:

2.

10. An isolated polypeptide comprising a variant of a fragment of SEQ ID NO:

2, wherein said fragment includes at least 500 consecutive amino acid residues of SEQ ID NO;

2.

11. The polypeptide according to claim 10, wherein the variant and said fragment have at least 91% sequence identity.

12. An antibody capable of binding to the amino acid sequence depicted in SEQ ID NO:

2 with a binding affinity of no less than 10⁵ M-1.

13. An MRCK2 detection kit comprising at least one of:

(a) the antibody of claim 12;

and

(b) a probe that is capable of hybridizing under stringent conditions to the nucleotide sequence of SEQ ID NO;

1 or the complement thereof.

16. A non-human animal, wherein at least one allele of a gene in the genome of said animal is functionally disrupted, and wherein said gene encodes a polypeptide that has at least 70% sequence identity to SEQ ID NO:

2.

17. A method for identifying an agent capable of binding to MRCK2 kinase, said method comprising:

contacting a candidate agent with a polypeptide comprising;

(a) an amino acid sequence recited in SEQ ID NO;

2,

(b) a fragment of SEQ ID NO;

2, or

(c) a variant of (a) or (b); and

detecting the binding between said candidate agent and said polypeptide.

18. A method for identifying an agent capable of modulating an activity of an MRCK2 kinase, comprising:

contacting a candidate agent with an polypeptide comprising;

(a) an amino acid sequence recited in SEQ ID NO;

2, or

(b) a biologically active portion of SEQ ID NO;

2; and

detecting a change in an activity of said polypeptide.

19. A pharmaceutical composition for preventing or treating MRCK2-related diseases, comprising a pharmaceutically acceptable carrier and an agent that modulates an MRCK2 activity or the MRCK2 gene expression.

20. A method for preventing or treating an MRCK2-related disease in a subject, said method comprising the step of:

introducing into the subject an effective amount of the pharmaceutical composition of claim 19.

21. A polynucleotide capable of inhibiting human MRCK2 gene expression by RNA interference.

22. The polynucleotide according to claim 21, comprising a siRNA sense strand or a siRNA antisense strand selected from Table 4.

23. A method, comprising introducing a polynucleotide of claim 21 into a cell which expresses human MRCK2 gene, thereby inhibiting the expression of said gene in said cell by RNA interference.