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Quantitative Dissection of the Allosteric and Sequence-Dependent Regulatory Genome in E. coli

Citation

Belliveau, Nathan Maurice (2018) Quantitative Dissection of the Allosteric and Sequence-Dependent Regulatory Genome in E. coli. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/Z9DN438T. https://resolver.caltech.edu/CaltechTHESIS:01052018-221609680

Abstract

Transcriptional regulation of gene expression is one of the most ubiquitous processes in biology. But while the catalog of bacterial genomes continues to expand rapidly, we remain ignorant about how almost all of the genes in these genomes are regulated. One of the ways genes are regulated is through external signals. To that end, we begin by presenting a general theory of allosteric transcriptional regulation using a statistical formulation of the Monod-Wyman-Changeux model, which we rigorously test using the ubiquitous simple repression motif in Escherichia coli. We then move to consider the consequence of the regulatory sequences themselves on gene expression. Here we apply a massively parallel reporter assay, Sort-Seq, to build models that describe the sequence-dependent binding energies of transcription factors and RNA polymerase to DNA. By coupling such models to our thermodynamic models of regulation, we construct a genotype to phenotype mapping that predicts gene expression as a function of regulatory sequence. We first demonstrate this approach in the context of the allosteric simple repression motif, and then show how it can be applied broadly across a bacterial genome, in conjunction with mass spectrometry, to uncover how genes are regulated.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:gene regulation, allostery, Allosteric regulation, LacI, lac operon, massively parallel reporter assay, Sort-Seq, DNA affinity chromatography, mass spectrometry, quantitative models
Degree Grantor:California Institute of Technology
Division:Biology and Biological Engineering
Major Option:Bioengineering
Research Advisor(s):
  • Phillips, Robert B.
Thesis Committee:
  • Phillips, Robert B. (chair)
  • Elowitz, Michael B.
  • Thomson, Matthew
  • Van Valen, David A.
Defense Date:14 December 2017
Record Number:CaltechTHESIS:01052018-221609680
Persistent URL:https://resolver.caltech.edu/CaltechTHESIS:01052018-221609680
DOI:10.7907/Z9DN438T
Related URLs:
URLURL TypeDescription
https://doi.org/10.1101/111013DOIbioRxiv preprint related to Chapter 2.
https://doi.org/10.1101/239335DOIbioRxiv preprint related to Chapter 4.
ORCID:
AuthorORCID
Belliveau, Nathan Maurice0000-0002-1536-1963
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:10623
Collection:CaltechTHESIS
Deposited By: Nathan Belliveau
Deposited On:17 Jan 2018 23:56
Last Modified:04 Oct 2019 00:19

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