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A detailed analysis of the DNA binding properties and the affinity purification of the Drosophila heat-shock transcription factor

Citation

Shuey, David J. (2002) A detailed analysis of the DNA binding properties and the affinity purification of the Drosophila heat-shock transcription factor. Dissertation (Ph.D.), California Institute of Technology. http://resolver.caltech.edu/CaltechTHESIS:01312012-161653671

Abstract

The heat shock response presents an extremely attractive model to study the regulation of gene transcription in eucaryotes. I have focused the bulk of my research efforts on investigating the molecular determinants of protein-DNA interaction exhibited by the heat-shock transcription factor (HSTF) of Drosophila. The specific "contacts" made by the HSTF upon binding to the heat-shock element (HSE) were exhaustively determined using a variety of chemical and enzymatic probes for a number of HSTF-HSE complexes formed at both hsp70 and hsp83 gene promoters. During the course of these studies it is demonstrated that the HSTF appears to polymerize in a sequence-specific and template-directed manner on each of these promoters, a novelty in this class of regulators. Evidence suggesting that DNA bending may occur during this HSTF-DNA association on the hsp70 promoter is also presented. This observation represented the first report of a eucaryotic transcriptional activator exhibiting this property.

Significantly, during the course of these studies two novel technologies were advanced; namely the gel-based contact point method and sequence-specific DNA affinity purification methodology. These technical advancements and the HSTF-HSE interaction results will be discussed in the body of the thesis in light of their relevance to both heat-shock and global gene expression mechanisms.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:Biochemistry and Molecular Biophysics
Degree Grantor:California Institute of Technology
Division:Chemistry and Chemical Engineering
Major Option:Biochemistry and Molecular Biophysics
Thesis Availability:Restricted to Caltech community only
Research Advisor(s):
  • Parker, Carl Stevens
Thesis Committee:
  • Attardi, Giuseppe
  • Wold, Barbara J.
  • Abelson, John N.
Defense Date:16 July 2001
Record Number:CaltechTHESIS:01312012-161653671
Persistent URL:http://resolver.caltech.edu/CaltechTHESIS:01312012-161653671
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:6800
Collection:CaltechTHESIS
Deposited By: John Wade
Deposited On:01 Feb 2012 15:43
Last Modified:26 Dec 2012 04:40

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