Abstract

Hsp90 is a ubiquitous heat shock protein that has been described mainly in terms of its response to stress an its association with steroid receptors and other proteins. Its function remains unknown. The long- term objective of these studies is to identify functional elements of hsp90. This will be approached through mutational mapping of hsp90 domains of interaction with other cellular components. A series of hsp90 deletion mutants has been prepared and these in a cell-free system. Two additional novel tests for hsp90 mutants will be developed in Aim 1. The first concerns a more general complex of hsp90 with two involving firefly luciferase renaturation. A third phase of mutant testing will be conducted in Aim 2. This will include tests on hsp90 dimerization, the binding and hydrolysis of ATP, and hsp90 binding to another associated protein, p59 or FKBP59.
In Aim 3, a new series of hsp90 mutants will be prepared and tested, based on the information gained from Aims 1 and 2. These mutants will be to obtain precise mutant that lack a specific interaction or function with a minimum of structural alteration in the molecule. These mutants will be used to describe separate and interrelated functional domains on hsp90 that are important to its biological activities.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK046249-02
Application #
2145421
Study Section
Biochemical Endocrinology Study Section (BCE)
Project Start
1993-12-01
Project End
1998-11-30
Budget Start
1994-12-01
Budget End
1995-11-30
Support Year
2
Fiscal Year
1995
Total Cost
Indirect Cost

  Institution

Name
Mayo Clinic, Rochester
Department
Type
DUNS #
City
Rochester
State
MN
Country
United States
Zip Code
55905

  Publications

Zhang, Zhongming; Sullivan, William; Felts, Sara J et al. (2010) Characterization of plant p23-like proteins for their co-chaperone activities. Cell Stress Chaperones 15:703-15
Moffatt, Nela S Cintron; Bruinsma, Elizabeth; Uhl, Cindy et al. (2008) Role of the cochaperone Tpr2 in Hsp90 chaperoning. Biochemistry 47:8203-13
Chadli, Ahmed; Bruinsma, Elizabeth S; Stensgard, Bridget et al. (2008) Analysis of Hsp90 cochaperone interactions reveals a novel mechanism for TPR protein recognition. Biochemistry 47:2850-7
Felts, Sara J; Karnitz, Larry M; Toft, David O (2007) Functioning of the Hsp90 machine in chaperoning checkpoint kinase I (Chk1) and the progesterone receptor (PR). Cell Stress Chaperones 12:353-63
Arlander, Sonnet J H; Felts, Sara J; Wagner, Jill M et al. (2006) Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones. J Biol Chem 281:2989-98
Huai, Qing; Wang, Huanchen; Liu, Yudong et al. (2005) Structures of the N-terminal and middle domains of E. coli Hsp90 and conformation changes upon ADP binding. Structure 13:579-90
Pratt, William B; Toft, David O (2003) Regulation of signaling protein function and trafficking by the hsp90/hsp70-based chaperone machinery. Exp Biol Med (Maywood) 228:111-33
Wang, Liewei; Sullivan, William; Toft, David et al. (2003) Thiopurine S-methyltransferase pharmacogenetics: chaperone protein association and allozyme degradation. Pharmacogenetics 13:555-64
Sullivan, William P; Owen, Barbara A L; Toft, David O (2002) The influence of ATP and p23 on the conformation of hsp90. J Biol Chem 277:45942-8
Hu, Jianming; Toft, David; Anselmo, Dana et al. (2002) In vitro reconstitution of functional hepadnavirus reverse transcriptase with cellular chaperone proteins. J Virol 76:269-79

Showing the most recent 10 out of 20 publications