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| Status |
Public on Feb 24, 2010 |
| Title |
Nicotinamide Starvation of Mycobacteria Defective for de novo NAD Biosynthesis |
| Platform organism |
Mycobacterium tuberculosis |
| Sample organisms |
Mycobacterium tuberculosis variant bovis; Mycobacterium tuberculosis |
| Experiment type |
Expression profiling by array
|
| Summary |
Mycobacteria can synthesize NAD+ using either the de novo biosynthesis pathway or the salvage pathway. The deletion of the three genes involved specifically in the NAD+ de novo biosynthesis pathway in the human pathogen Mycobacterium tuberculosis had no effect on the growth of the strain in vivo. In contrast, the same deletion in the bovine pathogen Mycobacterium bovis resulted in a strain that could not grow in vivo and could only grow in vitro with substantial nicotinamide supplmentation. This striking difference was attributed to the known defect in the nicotinamidase PncA of M. bovis, since introducing the M. tuberculosis pncA gene into the M. bovis strain defective for de novo NAD+ biosynthesis restored growth in vitro and in vivo. This study demonstrates that NAD+ starvation is a cidal event in mycobacteria and confirms that enzymes common to the de novo and salvage pathways may be good drug targets. We also propose that simultaneously targeting both the salvage and the de novo NAD+ biosynthesis pathways represents a potentially effective way to treat infection with tubercle bacilli. To characterize the lethality induced by nicotinamide starvation transcriptional profiling of the auxotrophs was performed. Triplicate 50 mL cultures of M. tuberculosis and M. bovis Delta nadABC mutants were grown in 7H9 OADC glycerol 0.05% tween broth in 500 mL roller bottles to an OD600nm= 0.1 in a roller incubator at 37°C. The cells were washed 1x in PBS and resuspended in 50 mL 7H9 OADC glycerol 0.05% tween broth with or without 20mg/L nicotinamide and returned to the incubator. After 7 days, cultures were harvested.
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| Overall design |
Three biological replicates of each of two species with one dye-flip each
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| Contributor(s) |
Weinrick B, Vilcheze C |
| Citation(s) |
20199601 |
| Submission date |
Nov 05, 2009 |
| Last update date |
Nov 12, 2013 |
| Contact name |
Brian Weinrick |
| Organization name |
Howard Hughes Medical Institute at Albert Einstein College of Medicine
|
| Department |
Microbiology & Immunology
|
| Lab |
Jacobs Lab, Price Center Room 569
|
| Street address |
1300 Morris Park Avenue
|
| City |
Bronx |
| State/province |
NY |
| ZIP/Postal code |
10461 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL5774 |
JCVI PFGRC Mycobacterium tuberculosis 22K v4 array designed primarily based on strain H37Rv |
|
| Samples (6)
|
| GSM468504 |
Nicotinamide Starve of Mycobacterium tuberculosis Defective for de novo NAD Biosysthesis Replicate 1 |
| GSM468505 |
Nicotinamide Starve of Mycobacterium tuberculosis Defective for de novo NAD Biosysthesis Replicate 2 |
| GSM468506 |
Nicotinamide Starve of Mycobacterium tuberculosis Defective for de novo NAD Biosysthesis Replicate 3 |
| GSM468507 |
Nicotinamide Starve of Mycobacterium bovis Defective for de novo NAD Biosysthesis Replicate 1 |
| GSM468526 |
Nicotinamide Starve of Mycobacterium bovis Defective for de novo NAD Biosysthesis Replicate 2 |
| GSM468527 |
Nicotinamide Starve of Mycobacterium bovis Defective for de novo NAD Biosysthesis Replicate 3 |
|
| Relations |
| BioProject |
PRJNA121323 |
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