Identifying mechanisms of regulation and signal integration of the heat shock response

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The heat shock response (HSR) is the cell's primary response to cytosolic misfolded protein. It is conserved across eukaryotes and is essential for cell survival. How cells detect misfolded protein and modulate their HSR levels is a poorly understood process. Additionally, manipulation of HSR levels has shown significant promise in treating diseases including cancer and neurodegeneration. Many mechanisms have been proposed for how misfolded protein could alter HSR activity. However, these regulatory mechanisms often lack evidence for if or when they are ever employed. Thus, for most proteotoxic stress conditions, if one or more of these proposed mechanisms is used to activate the HSR or if HSR activation occurs through an entirely different mechanism remains unknown. In this dissertation, I describe our work to quantify the contributions of known HSR activation pathways and discover new pathways of activating the HSR. We developed a method to quantify the most prevalent model of HSR regulation: inhibition of the HSR by the Hsp90 chaperone. We found some stressors for which this model can explain much of the HSR activation, and others for which it contributed very little, suggesting alternative mechanisms of HSR activation. Additionally, I describe the development of ReporterSeq, a high throughput screen to identify genes which regulate a reporter gene's expression under multiple conditions in a time-resolved manner. Using ReporterSeq, we measured the effect of full genome knockdowns on HSR activation across 15 stress conditions. From these data, we identified new general and stress-specific genetic regulators of the HSR. Combined, these data give us a more complete idea of the regulatory mechanisms of the HSR and how they are integrated together


Type of resource text
Form electronic resource; remote; computer; online resource
Extent 1 online resource
Place California
Place [Stanford, California]
Publisher [Stanford University]
Copyright date 2020; ©2020
Publication date 2020; 2020
Issuance monographic
Language English


Author Alford, Brian Douglas
Degree supervisor Brandman, Onn
Thesis advisor Brandman, Onn
Thesis advisor Krasnow, Mark, 1956-
Thesis advisor Rohatgi, Rajat
Degree committee member Krasnow, Mark, 1956-
Degree committee member Rohatgi, Rajat
Associated with Stanford University, Department of Biochemistry.


Genre Theses
Genre Text

Bibliographic information

Statement of responsibility Brian Douglas Alford
Note Submitted to the Department of Biochemistry
Thesis Thesis Ph.D. Stanford University 2020
Location electronic resource

Access conditions

© 2020 by Brian Douglas Alford
This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).

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