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A PUTATIVE DRUG EFFLUX PUMP IN MYCOBACTERIA:
EXAMINING THE PROMOTER STRUCTURE AND REGULATION OF THE
NOVEL PROTEIN MMPS-3
by
Alex Michael Kasman
A Thesis Presented to the
FACULTY OF THE USC GRADUATE SCHOOL
UNIVERSITY OF SOUTHERN CALIFORNIA
In Partial Fulfillment of the
Requirements for the Degree
MASTER OF SCIENCE
(EXPERIMENTAL AND MOLECULAR PATHOLOGY)
August 2011
Copyright 2011 Alex Michael Kasman
Object Description
| Title | A putative drug efflux pump in mycobacteria: examining promoter structure and regulation of the novel protein mmpS-3 |
| Author | Kasman, Alex Michael |
| Author email | akasman@gmail.com;kasman@usc.edu |
| Degree | Master of Science |
| Document type | Thesis |
| Degree program | Experimental and Molecular Pathology |
| School | Keck School of Medicine |
| Date defended/completed | 2011-05-25 |
| Date submitted | 2011-06-23 |
| Date approved | 2011-06-23 |
| Restricted until | 2011-12-23 |
| Date published | 2011-12-23 |
| Advisor (committee chair) | Ouellette, Andre |
| Advisor (committee member) |
Hofman, Florence McMillan, Minnie Nash, Kevin |
| Abstract | Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), caused nearly 2 million deaths in 2009 while one third of the world population is likely infected with the bacillus. Efforts to control this deadly pathogen have increased greatly and been successful, but growing antibiotic resistance presents an important roadblock to elimination of TB as a public health concern. While genomic mutations leading to resistance to chemotherapeutic treatment in mycobacteria have been characterized and observed clinically, drug efflux pumps have received little attention and likely present a future threat. Our lab has identified a putative drug efflux pump, Mmps-3, in mycobacteria that when knocked-out increases susceptibility to macrolide antimicrobial agents. However, beyond this limited knowledge little is known about this gene, thus the aim of this study was to characterize the promoter structure and regulation of mmpS-3. Promoter mapping has revealed the mmpS-3 promoter region in both M. bovis BCG and M. smegmatis mc²155 while truncation mutants have begun to identify the minimal promoter region in M. smegmatis mc²155. Initial screening using GFP as a reporter gene suggests that the mmpS-3 promoter is constitutively active, but at relatively low levels both in the presence and absence of ampicillin, erythromycin, isoniazid, and rifampicin. Expression analysis showed that MmpS-3, as well as sigma factor A (sigA) expression significantly increased when sigma factor B (sigB) expression was induced by heat shock and in trans over expression. This study has begun to analyze the promoter structure and regulation of the novel protein MmpS-3 and has demonstrated that sigB regulates mmpS-3 either directly or indirectly. |
| Keyword | mycobacteria; efflux; promoter; tuberculosis |
| 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 | Kasman, Alex Michael |
| 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_Volume71/etd-KasmanAlex-32.pdf |
Description
| Title | Page 1 |
| Full text | A PUTATIVE DRUG EFFLUX PUMP IN MYCOBACTERIA: EXAMINING THE PROMOTER STRUCTURE AND REGULATION OF THE NOVEL PROTEIN MMPS-3 by Alex Michael Kasman A Thesis Presented to the FACULTY OF THE USC GRADUATE SCHOOL UNIVERSITY OF SOUTHERN CALIFORNIA In Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE (EXPERIMENTAL AND MOLECULAR PATHOLOGY) August 2011 Copyright 2011 Alex Michael Kasman |
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