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THE EFFECTS OF OVEREXPRESSION OF MTA: A PUTATIVE CELL
MEMBRANE PROTEIN IN MYCOBACTERIA
by
Emily Chau
____________________________________________________________________
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 Emily Chau
Object Description
| Title | The effects of overexpression of MtA: a putative cell membrane protein in mycobacteria |
| Author | Chau, Emily Man-Shan |
| Author email | emilymchau@gmail.com;emilymch@usc.edu |
| Degree | Master of Science |
| Document type | Thesis |
| Degree program | Experimental and Molecular Pathology |
| School | Keck School of Medicine |
| Date defended/completed | 2011-05-10 |
| Date submitted | 2011-07-18 |
| Date approved | 2011-07-18 |
| Restricted until | 2011-07-18 |
| Date published | 2011-07-18 |
| Advisor (committee chair) | Taylor, Clive R. |
| Advisor (committee member) |
Nash, Kevin A. Ouellette, Andre J. McMillan, Minnie |
| Abstract | Nontuberculosis mycobacterial infections are on a rise and are extremely difficult to treat. It is important to examine their mechanisms of reducing antimicrobial agents by their normal cellular functions so we can develop therapeutic strategies to counter this resistance to antimicrobial agents. Based on previous studies on knockout of mtA gene in Mycobacterium smegmatis, we hypothesize that the proteins encoded by the mtA-mtB operon are components of a transmembrane transport system that can efflux macrolide antimicrobial agents. It was found that overexpression of mtA-mtB operon in particular mtA is extremely detrimental to the organism. The only viable organisms that were recovered were ones with mutations in the mtA gene of the inserted plasmid after a short period of overexpression. However, we were able to restore the loss of function caused by the knockout of mtA in the organism through the use of an acetamidase promoter that is induced by acetamide. This acetamidase promoter at baseline is expressing the mtA-mtB operon and making proteins; thus, M. smegmatis’s (with knockout of mtA) susceptibility to erythromycin was lessened. We have not been successful in directly confirming proteins made from the mtA-mtB operon are indeed membrane proteins, but the data we have collected do suggest that these proteins are membrane proteins and they play a role in intrinsic resistance to macrolides. Therefore, understanding the normal cellular functions of this potential efflux pump and why overexpression is detrimental can help in developing new therapeutic drug regiments against mycobacteria. |
| Keyword | membrane Protein overexpression; M. smegmatis; mycobacteria |
| 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 |
| Contributing entity | University of Southern California |
| Rights | Chau, Emily Man-Shan |
| Physical access | 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@dots.usc.edu |
| Archival file | uscthesesreloadpub_Volume71/etd-ChauEmilyM-109-0.pdf |
Description
| Title | Page 1 |
| Contributing entity | University of Southern California |
| Repository email | cisadmin@dots.usc.edu |
| Full text | THE EFFECTS OF OVEREXPRESSION OF MTA: A PUTATIVE CELL MEMBRANE PROTEIN IN MYCOBACTERIA by Emily Chau ____________________________________________________________________ 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 Emily Chau |
