The heat shock transcription factor (HSF) is a transcriptional regulator whose response to heat shock and other stresses is to activate the expression of the heat shock proteins, a set of proteins that have critical constitutive and protective functions. Heat shock proteins also play multiple roles in several disease states, including cancer, cardiovascular problems, and neurodegenerative conditions involving protein aggregation. The ability to control the expression of heat shock proteins under different conditions could have profound effects on these disease states. Therefore, it has become important to understand how the activity of HSF is regulated, so that we can learn how to control its activity. HSF has a low level of activity prior to stress and acquires a high level of transcriptional activity after stress. The modulation of HSF's activity likely involves other regulatory proteins, post-translational modifications, and cellular localization. Of interest to this proposal is the fact that mutations and/or deletions within HSF can cause it to become constitutively active. Therefore, it is the integrity of the HSF structure that appears to be most critical for regulating its activity.
The specific aims of this proposal focus on the DNA-binding and trimerization domains, two domains for which much is known about the structure and function. Previous structural and genetic studies will guide the choice of HSF mutants to use to test hypotheses as to whether charge, surface topology, thermal stability, and/or overall structure are critical for the normal regulation of HSF's transcriptional activity. The ability of these mutant HSFs to differentially activate different heat shock promoters and to be differentially induced by different stresses will be also analyzed. These studies will deepen our understanding of how the structure of HSF relates to its activity and will help lead to the long term goal of being able to manipulate HSF's function in order to take advantage of its protective role.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM044086-13
Application #
6547060
Study Section
Biochemistry Study Section (BIO)
Program Officer
Wehrle, Janna P
Project Start
1990-04-01
Project End
2006-06-30
Budget Start
2002-07-01
Budget End
2003-06-30
Support Year
13
Fiscal Year
2002
Total Cost
$330,936
Indirect Cost
Name
University of Pennsylvania
Department
Biochemistry
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Dashnau, Jennifer L; Conlin, Laura K; Nelson, Hillary C M et al. (2008) Water structure in vitro and within Saccharomyces cerevisiae yeast cells under conditions of heat shock. Biochim Biophys Acta 1780:41-50
Conlin, Laura K; Nelson, Hillary C M (2007) The natural osmolyte trehalose is a positive regulator of the heat-induced activity of yeast heat shock transcription factor. Mol Cell Biol 27:1505-15
Zhao, Xiaoching; Shi, Hua; Sevilimedu, Aarti et al. (2006) An RNA aptamer that interferes with the DNA binding of the HSF transcription activator. Nucleic Acids Res 34:3755-61
Eastmond, Dawn L; Nelson, Hillary C M (2006) Genome-wide analysis reveals new roles for the activation domains of the Saccharomyces cerevisiae heat shock transcription factor (Hsf1) during the transient heat shock response. J Biol Chem 281:32909-21
Ferguson, Scott B; Anderson, Erik S; Harshaw, Robyn B et al. (2005) Protein kinase A regulates constitutive expression of small heat-shock genes in an Msn2/4p-independent and Hsf1p-dependent manner in Saccharomyces cerevisiae. Genetics 169:1203-14
Bulman, Amanda L; Nelson, Hillary C M (2005) Role of trehalose and heat in the structure of the C-terminal activation domain of the heat shock transcription factor. Proteins 58:826-35
Cicero, M P; Hubl, S T; Harrison, C J et al. (2001) The wing in yeast heat shock transcription factor (HSF) DNA-binding domain is required for full activity. Nucleic Acids Res 29:1715-23
Littlefield, O; Nelson, H C (2001) Crystal packing interaction that blocks crystallization of a site-specific DNA binding protein-DNA complex. Proteins 45:219-28
Bulman, A L; Hubl, S T; Nelson, H C (2001) The DNA-binding domain of yeast heat shock transcription factor independently regulates both the N- and C-terminal activation domains. J Biol Chem 276:40254-62
Hardy, J A; Nelson, H C (2000) Proline in alpha-helical kink is required for folding kinetics but not for kinked structure, function, or stability of heat shock transcription factor. Protein Sci 9:2128-41

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