DOT1 histone methyltransferases as a target for identifying therapeutic agents for leukemia

• US 7,442,685 B2
• Filed: 06/14/2004
• Issued: 10/28/2008
• Est. Priority Date: 06/13/2003
• Status: Expired due to Fees

• Current Assignee: University of North Carolina At Chapel Hill (University of North Carolina System)
• Original Assignee: University of North Carolina At Chapel Hill (University of North Carolina System)
• Inventors: Zhang, Yi, Okada, Yuki, Xu, Guoliang, Feng, Qin
• Primary Examiner(s): Fetterolf; Brandon J
• Application Number:
  US10/866,908
• Publication Number:
  US 20050048634A1
• Time in Patent Office: 1,597 Days
• Field of Search: 514/12
• US Class Current: 514/1.1
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12N 9/1007 Methyltransferases (general) (2.1.1.)
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/118 Prognosis of disease development
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/154 Methylation markers
• CPC Class Codes: G01N 2440/12 alkylation, e.g. methylation, (iso-)prenylation, farnesylation
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

The present invention provides polypeptides with histone H3 lysine 79 methyltransferase activity as well as nucleic acids encoding the same. Also provided are methods of using the polypeptides and nucleic acids of the invention in screening assays to identify compounds of interest. Further provided are diagnostic methods for leukemia and prognostic methods to predict the course of the disease in a subject.

Claims

1. An isolated DOT1L polypeptide comprising the amino acid sequence of SEQ ID NO:

2.

2. The isolated DOT1L polypeptide of claim 1 consisting of the amino acid sequence of SEQ ID NO:

2.

3. A fusion protein comprising the DOT1L polypeptide of claim 1.

4. An isolated functional fragment of SEQ ID NO:

2 comprising at least 400 amino acids of the amino acid sequence of SEQ ID NO;

2 or an amino acid sequence having at least 95% amino acid sequence similarity thereto, wherein said functional fragment comprises a DOT1L histone methyltransferase catalytic domain and has H3-K79 methyltransferase activity, further wherein said domain comprises amino acids 157 to 270 of SEQ ID NO;

2 or an amino acid sequence having at least 95% amino acid sequence similarity thereto.

5. The isolated functional fragment of claim 4, wherein the catalytic domain of the functional fragment further comprises amino acids 121-306 of SEQ ID NO:

2 or an amino acid sequence having at least 95% amino acid sequence similarity thereto.

6. The isolated functional fragment of claim 4, further comprising amino acids 390 to 407 of SEQ ID NO:

2 or an amino acid sequence having at least 95% amino acid sequence similarity thereto.

7. The isolated functional fragment of claim 4 further comprising a leucine rich region that binds AF10.

8. The isolated functional fragment of claim 7, wherein the leucine rich region that binds AF10 comprises amino acids 500 to 630 of SEQ ID NO:

2 or an amino acid sequence having at least 95% amino acid sequence similarity thereto.

9. The isolated functional fragment of claim 4 further comprising a nuclear export signal.

10. The isolated functional fragment of claim 9, wherein the nuclear export signal comprises an amino acid sequence selected from the group consisting of:

(i) amino acids 482 to 496 of SEQ ID NO;

2 or an amino acid sequence having at least 95% amino acid sequence similarity thereto;

(ii) amino acids 600 to 635 of SEQ ID NO;

2 or an amino acid sequence having at least 95% amino acid sequence similarity thereto;

(iii) amino acid 636 to 650 of SEQ ID NO;

2 or an amino acid sequence having at least 95% amino acid sequence similarity thereto; and

(iv) a combination of any of (i) to (iii) above.

11. The isolated functional fragment of claim 9, wherein the nuclear export signal comprises amino acids 472 to 767 of SEQ ID NO:

2 or an amino acid sequence having at least 95% amino acid sequence similarity thereto.

12. A fusion protein comprising the isolated functional fragment of claim 4.