Histone deacetylases, and uses related thereto

• US 6,777,217 B1
• Filed: 03/26/1996
• Issued: 08/17/2004
• Est. Priority Date: 03/26/1996
• Status: Expired due to Fees

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Schreiber, Stuart L., Hassig, Christian A., Jamison, Timothy F., Taunton, Jack
• Primary Examiner(s): HUTSON, RICHARD G
• Application Number:
  US08/624,735
• Time in Patent Office: 3,066 Days
• Field of Search: 435/194, 435/252.3, 435/325, 435/320.1, 935/22, 935/52, 935/69-72, 935/76-78, 536/23.1, 536/23.2
• US Class Current: 435/194
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 31/165 having aromatic rings, e.g. colchicine, atenolol, progabide
• CPC Class Codes: A61K 31/407 condensed with other heterocyclic ring systems, e.g. ketorolac, physostigmine
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 14/4703 Inhibitors; Suppressors
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 5/126 Tetrapeptides
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)
• CPC Class Codes: C12N 9/80 acting on amide bonds in linear amides (3.5.1)

Abstract

The present invention concerns the discovery that proteins encoded by a family of genes, termed here HDx-related genes, which are involved in the control of chromatin structure and, thus in transcription and translation. The present invention makes available compositions and methods that can be utilized, for example to control cell proliferation and differentiation in vitro and in vivo.

Claims

1. An isolated nucleic acid encoding a histone deacetylase gene family (HDx) polypeptide having a histone deacetylase activity, which HDx polypeptide comprises a ν

motif represented in the general formula

DIAX₁NWAGGLHHAKKX₂EASGFCYVNDIVX₃X₄ILELLKYHX₅RVLYLDIDIHHGDGX₆EA FYX₇TDRVMTVSF (SEQ ID NO;

     13) and a χ

motif represented in CVEX₁VKX₂FNX₃P-X₄LX₅LGGGGYTX₆RNVARCWTYET (SEQ ID NO;

     16), and which nucleic acid hybridizes under stringent conditions of 6.0×

SSC at 65°

C. to the nucleotide sequence designated in SEQ ID NO;

1.

3. The nucleic acid of claim 1, which HDx polypeptide has a molecular weight in the range of 45-70 Kd.

4. The nucleic acid of claim 1, which HDx polypeptide is a fusion protein.

5. The nucleic acid of claim 4, wherein said fusion protein includes, as a second polypeptide sequence, a polypeptide which functions as a detectable label for detecting the presence of said fusion protein or as a matrix-binding domain for immobilizing said fusion protein.

6. The nucleic acid of claim 1, further comprising a transcriptional regulatory sequence operably linked to said nucleotide sequence so as to render said nucleic acid suitable for use as an expression vector.

7. An expression vector, capable of replicating in at least one of a prokaryotic cell and eukaryotic cell, comprising the nucleic acid of claim 6.

8. A host cell transfected with the expression vector of claim 7 and expressing said recombinant polypeptide.

9. A method of producing a recombinant HDx polypeptide comprising culturing the cell of claim 8 in a cell culture medium to express said recombinant polypeptide and isolating said recombinant polypeptide from said cell culture.

10. A recombinant transfection system, comprising:

(i) a gene construct including the nucleic acid of claim 1 and operably linked to a transcriptional regulatory sequence for causing expression of said HDx polypeptide in eukaryotic cells; and

(ii) a gene delivery composition for delivering said gene construct to a cell and causing the cell to be transfected with said gene construct.

11. The recombinant transfection system of claim 10, wherein the gene delivery composition is selected from the group consisting of a recombinant viral particle, a liposome, and a poly-cationic nucleic acid binding agent.

2. An isolated nucleic acid encoding an HDx polypeptide sequence or fragment thereof, which can deacetylate an acetylated histone substrate, and which polypeptide comprises the polypeptide sequence designated in SEQ ID NO:

5.