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Investigating the E. Coli DNA Polymerase V Mutasome
by
Meghna Patel
———————————
A Dissertation Presented to the
FACULTY OF THE USC GRADUATE SCHOOL
UNIVERSITY OF SOUTHERN CALIFORNIA
In Partial Fulfillment of the
Requirements for the Degree
DOCTOR OF PHILOSOPHY
MOLECULAR BIOLOGY
May 2013
Copyright 2013 Meghna Patel
Object Description
| Title | Investigating the E. Coli DNA polymerase V mutasome |
| Author | Patel, Meghna |
| Author email | meghnapa@usc.edu;meghnayadav@gmail.com |
| Degree | Doctor of Philosophy |
| Document type | Dissertation |
| Degree program | Molecular Biology |
| School | College of Letters, Arts And Sciences |
| Date defended/completed | 2012-10-05 |
| Date submitted | 2013-01-29 |
| Date approved | 2013-01-29 |
| Restricted until | 2013-01-29 |
| Date published | 2013-01-29 |
| Advisor (committee chair) | Goodman, Myron F. |
| Advisor (committee member) |
Pinaud, Fabien Reisler, Hanna |
| Abstract | In Escherichia Coli, cell survival and genomic stability after UV radiation or other DNA damaging agents depends on repair mechanisms induced as part of the SOS response. The SOS response involves upregulation of over 40 genes, which are normally repressed by the LexA protein under non-damage conditions. The early phase of the SOS response is centered around error-free DNA repair mechanisms. These include base excision repair (BER), nucleotide excision repair (NER), and recombinational repair. If these mechanisms are insufficient to surmount the replicational impedance brought about by DNA damage, the bacterial cell advances into the later part of the SOS response. This later phase of the response helps ensure cell survival albeit at the cost of faithful DNA repair. Known as SOS mutagenesis, this phase is characterized by elevated mutation levels, which are a direct result of error-prone DNA replication. SOS mutagenesis is the result of the low fidelity DNA polymerase V. DNA polymerase V (Pol V) is a heterotrimeric protein composed of the catalytically active UmuC and a dimer of UmuD'. The heterotrimer is essentially inactive, however, and must strip a RecA monomer and an ATP cofactor from the 3' tip of a RecA-DNA nucleoprotein filament to form an active Pol V mutasome composed of UmuC:UmuD'₂:RecA:ATP. This dissertation focuses on investigating the Pol V mutasome including deducing protein-protein interactions in the active, inactive, and de-activated states of the mutasome, detecting a conformational change in the different activity states of the mutasome, visualizing the SOS response in live cells, and attempting to ascertain the crystal structure of Pol V. |
| Keyword | translesion DNA synthesis; DNA Polymerase V; error-prone DNA repair; UV mutagenesis |
| Language | English |
| Part of collection | University of Southern California dissertations and theses |
| Publisher (of the original version) | University of Southern California |
| Place of publication (of the original version) | Los Angeles, California |
| Publisher (of the digital version) | University of Southern California. Libraries |
| Provenance | Electronically uploaded by the author |
| Type | texts |
| Legacy record ID | usctheses-m |
| Rights | Patel, Meghna |
| Access conditions | The author retains rights to his/her dissertation, thesis or other graduate work according to U.S. copyright law. Electronic access is being provided by the USC Libraries in agreement with the author, as the original true and official version of the work, but does not grant the reader permission to use the work if the desired use is covered by copyright. It is the author, as rights holder, who must provide use permission if such use is covered by copyright. The original signature page accompanying the original submission of the work to the USC Libraries is retained by the USC Libraries and a copy of it may be obtained by authorized requesters contacting the repository e-mail address given. |
| Repository name | University of Southern California Digital Library |
| Repository address | USC Digital Library, University of Southern California, University Park Campus MC 7002, 106 University Village, Los Angeles, California 90089-7002, USA |
| Repository email | cisadmin@usc.edu |
| Archival file | uscthesesreloadpub_Volume6/etd-PatelMeghn-1414.pdf |
Description
| Title | Page 1 |
| Full text | 1 Investigating the E. Coli DNA Polymerase V Mutasome by Meghna Patel ——————————— A Dissertation Presented to the FACULTY OF THE USC GRADUATE SCHOOL UNIVERSITY OF SOUTHERN CALIFORNIA In Partial Fulfillment of the Requirements for the Degree DOCTOR OF PHILOSOPHY MOLECULAR BIOLOGY May 2013 Copyright 2013 Meghna Patel |
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