Structural and Functional Characterization of IclR Transcription Regulators

This work is a part of a large project in our laboratory that is aimed toward characterization of prokaryotic transcription regulators from different families and their interactions with small-molecule effectors. My study was focused of IclR family of transcriprion regulators, specifically on its fo...

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Bibliographic Details
Main Author: Ezersky, Alexandra
Other Authors: Edwards, Aled M.
Language:en_ca
Published: 2009
Subjects:
Online Access:http://hdl.handle.net/1807/18291
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spelling ndltd-LACETR-oai-collectionscanada.gc.ca-OTU.1807-182912013-11-02T03:43:30ZStructural and Functional Characterization of IclR Transcription RegulatorsEzersky, AlexandraTranscription regulatorsX-ray crystallography0760This work is a part of a large project in our laboratory that is aimed toward characterization of prokaryotic transcription regulators from different families and their interactions with small-molecule effectors. My study was focused of IclR family of transcriprion regulators, specifically on its founding member Isocytrate Lyase Regulator (IclR) from E.coli and AllR regulator from E.coli, which share 42% sequence identity with IclR. I used a combination of biophysical, biochemical and structural biology techniques to explore the mechanisms by which IclR and AllR interact with their effectors. I performed site-directed mutagenesis experiments in order to research the role of individual amino acids in interaction of AllR regulator with its previously identified effector glyoxylate and to test whether oligomerization plays a role in effector-induced signal transduction by AllR. Using differential light scattering, which allows high-throughput screening of small molecules for thermostabilization of proteins, I identified potential effctors for the IclR regulator. The physiological relevance of these candidate molecules was tested in-vitro and in-vivo and their interaction with IclR was characterized by Isothermal Titration Calorimetry and X-ray Crystallography.Edwards, Aled M.2009-112010-01-15T19:17:22ZNO_RESTRICTION2010-01-15T19:17:22Z2010-01-15T19:17:22ZThesishttp://hdl.handle.net/1807/18291en_ca
collection NDLTD
language en_ca
sources NDLTD
topic Transcription regulators
X-ray crystallography
0760
spellingShingle Transcription regulators
X-ray crystallography
0760
Ezersky, Alexandra
Structural and Functional Characterization of IclR Transcription Regulators
description This work is a part of a large project in our laboratory that is aimed toward characterization of prokaryotic transcription regulators from different families and their interactions with small-molecule effectors. My study was focused of IclR family of transcriprion regulators, specifically on its founding member Isocytrate Lyase Regulator (IclR) from E.coli and AllR regulator from E.coli, which share 42% sequence identity with IclR. I used a combination of biophysical, biochemical and structural biology techniques to explore the mechanisms by which IclR and AllR interact with their effectors. I performed site-directed mutagenesis experiments in order to research the role of individual amino acids in interaction of AllR regulator with its previously identified effector glyoxylate and to test whether oligomerization plays a role in effector-induced signal transduction by AllR. Using differential light scattering, which allows high-throughput screening of small molecules for thermostabilization of proteins, I identified potential effctors for the IclR regulator. The physiological relevance of these candidate molecules was tested in-vitro and in-vivo and their interaction with IclR was characterized by Isothermal Titration Calorimetry and X-ray Crystallography.
author2 Edwards, Aled M.
author_facet Edwards, Aled M.
Ezersky, Alexandra
author Ezersky, Alexandra
author_sort Ezersky, Alexandra
title Structural and Functional Characterization of IclR Transcription Regulators
title_short Structural and Functional Characterization of IclR Transcription Regulators
title_full Structural and Functional Characterization of IclR Transcription Regulators
title_fullStr Structural and Functional Characterization of IclR Transcription Regulators
title_full_unstemmed Structural and Functional Characterization of IclR Transcription Regulators
title_sort structural and functional characterization of iclr transcription regulators
publishDate 2009
url http://hdl.handle.net/1807/18291
work_keys_str_mv AT ezerskyalexandra structuralandfunctionalcharacterizationoficlrtranscriptionregulators
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