Mycobacterium tuberculosis is responsible for more human deaths than any other single infectious agent and imposes a considerable social and economic toll on a global scale. Adult pulmonary tuberculosis is notoriously difficult to control, requiring extended treatments with multiple antibiotics. Lack of compliance with these regimens has encouraged the appearance of multiple drug-resistant strains of M. tuberculosis. ? ? Effective control of tuberculosis requires better diagnostic methods, more efficacious vaccines, and better anti-mycobacterial drugs, particularly for treatments of latent and multi-drug resistant infections. Recent advances in mycobacterial genetics and the availability of several whole mycobacterial genome sequences brings considerable promise to the idea that elucidating the fundamental tenets of mycobacterial physiology, metabolism, genetics and structure will lead to novel strategies for controlling mycobacterial infections. Currently, we know little about the molecular basis of the fundamental aspects of such central questions as to why these bacteria grow so slowly, how is their unique cell wall synthesized, and how do mycobacterial cells communicate. Answering these central questions is important to understanding the molecular basis of mycobacterial pathogenesis and how we might intervene in it. ? ? Bacteriophages are powerful tools for understanding their bacterial hosts, and this is certainly true for the mycobacteriophages. Mycobacteriophages are partners in an extremely intimate relationship with their hosts; they are not only dependent on the host metabolic machinery for their replication and assembly, but they must also gain entry through a complex and hostile mycobacterial cell wall, and find ways to exit the cell once it is no longer of any use to them. Furthermore, mycobacteriophages may not kill their hosts at all but integrate their DNA into the host chromosome and co-exist peacefully. ? ? This project focuses on examining ways in which mycobacteriophages mimic the cellular signaling pathways of the mycobacteria. We have discovered that mycobacteriophage Bxbl integrates into the groEL1 gene, one of two groEL genes in the mycobacteria. Interruption of the groEL1 gene leads to the loss of biofilm maturation, apparently due to the inability to assemble the FAS-II enzyme complex for mycolate synthesis. We propose to elucidate the specific role of GroEL1 in biofilm formation and mycolic acid synthesis, and to determine its role in the pathogenesis of Mycobacterium tuberculosis. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI064494-02
Application #
7244389
Study Section
Special Emphasis Panel (ZRG1-HIBP-H (01))
Program Officer
Lacourciere, Karen A
Project Start
2006-06-15
Project End
2011-05-31
Budget Start
2007-06-01
Budget End
2008-05-31
Support Year
2
Fiscal Year
2007
Total Cost
$511,967
Indirect Cost
Name
University of Pittsburgh
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Ojha, Anil K; Jacobs Jr, William R; Hatfull, Graham F (2015) Genetic dissection of mycobacterial biofilms. Methods Mol Biol 1285:215-26
Sambandan, Dhinakaran; Dao, Dee N; Weinrick, Brian C et al. (2013) Keto-mycolic acid-dependent pellicle formation confers tolerance to drug-sensitive Mycobacterium tuberculosis. MBio 4:e00222-13
Ojha, Anil K; Trivelli, Xavier; Guerardel, Yann et al. (2010) Enzymatic hydrolysis of trehalose dimycolate releases free mycolic acids during mycobacterial growth in biofilms. J Biol Chem 285:17380-9
Traag, Bjorn A; Driks, Adam; Stragier, Patrick et al. (2010) Do mycobacteria produce endospores? Proc Natl Acad Sci U S A 107:878-81
Payne, Kimberly; Sun, Qingan; Sacchettini, James et al. (2009) Mycobacteriophage Lysin B is a novel mycolylarabinogalactan esterase. Mol Microbiol 73:367-81
Ojha, Anil; Hatfull, Graham F (2007) The role of iron in Mycobacterium smegmatis biofilm formation: the exochelin siderophore is essential in limiting iron conditions for biofilm formation but not for planktonic growth. Mol Microbiol 66:468-83