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
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|
|Defense Date:||16 July 2001|
|Default Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||John Wade|
|Deposited On:||01 Feb 2012 15:43|
|Last Modified:||26 Dec 2012 04:40|
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