BH3 interacting domain death agonist

• US 5,955,593 A
• Filed: 09/09/1996
• Issued: 09/21/1999
• Est. Priority Date: 09/09/1996
• Status: Expired due to Term

• Current Assignee: Washington University In St Louis
• Original Assignee: Washington University In St Louis
• Inventors: Korsmeyer, Stanley J.
• Primary Examiner(s): Hutzell, Paula K.
• Assistant Examiner(s): Worrall, Timothy A.
• Application Number:
  US08/706,741
• Time in Patent Office: 1,107 Days
• Field of Search: 514/44, 536/23.1
• US Class Current: 536/23.5
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 14/4747 Apoptosis related proteins
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

A novel death agonist, BID, which is related to the BCL-2 family of apoptosis regulating proteins only in the conserved BH3 domain, is provided along with encoding polynucleotides. BID polypeptides and derivatives thereof are useful in modulating apoptosis. Polynucleotides encoding BID polypeptides and derivatives thereof are useful as templates for recombinantly expressing BID polypeptides, as probes for detecting endogenously expressed BID polynucleotides, and in gene therapy for treating or preventing disease. Methods for modulating inappropriate expression of BID using BID antisense molecules or anti-BID antibodies are also provided. In addition, screening assays for identifying agents which modulate binding between BID and other BCL-2 family members are provided.

Claims

1. An isolated and purified polynucleotide or a polynucleotide complementary thereto, wherein the polynucleotide comprises a nucleotide sequence which encodes a mammalian bcl-homology-domain-3-interacting-domain-death-agonist (BIH3-interacting-domain-death-agonist (BID)) polypeptide which (a) lacks a carboxyl terminal signal-anchor sequence characteristic of membrane bound BCL-2 family members, (b) lacks bcl-homology-domains 1 and 2, (c) has a BH3 domain, (d) heterodimerizes with BAX, BCl,-2 and BCL-X_L, and (e) has at least 85% sequence conservation with SEQ ID NO:

4, SEQ ID NO;

5, or SEQ ID NO;

6.

2. The polynucleotide or polynucleotide fully complementary thereto of claim 1 wherein said polynucleotide comprises a nucleotide sequence as set forth in SEQ ID NO:

1.

3. The polynucleotide or polynucleotide fully complementary thereto of claim 1 wherein said polynucleotide comprises a nucleotide sequence as set forth in SEQ ID NO:

2 or SEQ ID NO;

3.

10. A vector comprising expression regulatory elements operably linked to the polynucleotide or polynucleotide fully complementary thereto of claim 1.

11. A host cell transformed with the vector of claim 10.

14. A composition comprising:

(a) a fusion gene comprising the polynucleotide of claim 1 fused in frame to a polynucleotide encoding a DNA-binding domain of a transcriptional activator protein;

(b) a fusion gene comprising a sequence encoding BAX, BCL-2 or BCL-X_L fused in frame to a polynucleotide encoding a DNA-binding domain of the transcriptional activator protein of (a); and

(c) a reporter gene functionally linked to a transcriptional regulatory element whose transcriptional activity is dependent upon the amount of a heterodimer or peptides encoded by fusion genes of (a) and (b).

15. The composition of claim 14, wherein the transcriptional activator protein is the yeast GAL4 transcription protein.

16. The composition of claim 15, wherein the reporter gene is β

-galactosidase.

4. An isolated and purified polynucleotide or polynucleotide fully complementary thereto, wherein the polynucleotide comprises a nucleotide sequence encoding a fusion protein comprising SEQ ID NO:

54 and at least 10 consecutive amino acids of SEQ ID NO;

4, SEQ ID NO;

5, or SEQ ID NO;

6.

5. The polynucleotide or polynucleotide complementary thereto of claim 4, wherein the polynucleotide encodes a fusion protein having an amino acid sequence as set forth in SEQ ID NO:

56.

6. An isolated and purified polynucleotide comprising a nucleotide sequence which encodes a human bcl-homology-domain-3-interacting-domain-death-agonist (BH3-interacting-domain-death-agonist(BID)) polypeptide or a polynucleotide fully complementary thereto, wherein the BID polypeptide comprises an amino acid sequence which has at least 85 percent conservation with the amino acid sequence set forth in SEQ ID NO:

4, SEQ ID NO;

5, or SEQ ID NO;

6, and wherein the complementary polynucleotide specifically hybridizes to SEQ ID NO;

1, SEQ ID NO;

2 or SEQ ID NO;

3.

7. The polynucleotide or polynucleotide fully complementary thereto of claim 6, wherein the BID polypeptide comprises an amino acid sequence which is at least 85 percent identical to the amino acid sequence set forth in SEQ ID NO:

4 and wherein the complementary polynucleotide specifically hybridizes to SEQ ID NO;

1.

8. The polynucleotide or polynucleotide fully complementary thereto of claim 7, wherein the BID polypeptide comprises an amino acid sequence which has at least 95 percent conservation with the amino acid sequence set forth in SEQ ID NO:

4 and the complementary polynucleotide specifically hybridizes to SEQ ID NO;

1.

9. The polynucleotide or polynucleotide fully complementary thereto of claim 8, wherein the BID polypeptide comprises an amino acid sequence which is at least 95 percent identical to the amino acid sequence set forth in SEQ ID NO:

4 and the complementary polynucleotide specifically hybridizes to SEQ ID NO;

1.

12. A vector comprising expression regulatory elements operably linked to the polynucleotide or polynucleotide fully complementary thereto of claim 6.

13. A host cell transforced with the vector of claim 12.