Transcriptional Adaptation of Mycobacterium tuberculosis within Macrophages: Insights into the Phagosomal Environment.
Little is known about the biochemical environment in phagosomes harboring an infectious agent. To assess the state of this organelle we captured the transcriptional responses of Mycobacterium tuberculosis (MTB) in macrophages from wild-type and nitric oxide (NO) synthase 2-deficient mice before and after immunologic activation. The intraphagosomal transcriptome was compared with the transcriptome of MTB in standard broth culture and during growth in diverse conditions designed to simulate features of the phagosomal environment. Genes expressed differentially as a consequence of intraphagosomal residence included an interferon gamma- and NO-induced response that intensifies an iron-scavenging program, converts the microbe from aerobic to anaerobic respiration, and induces a dormancy regulon. Induction of genes involved in the activation and beta-oxidation of fatty acids indicated that fatty acids furnish carbon and energy. Induction of sigmaE-dependent, sodium dodecyl sulfate-regulated genes and genes involved in mycolic acid modification pointed to damage and repair of the cell envelope. Sentinel genes within the intraphagosomal transcriptome were induced similarly by MTB in the lungs of mice. The microbial transcriptome thus served as a bioprobe of the MTB phagosomal environment, showing it to be nitrosative, oxidative, functionally hypoxic, carbohydrate poor, and capable of perturbing the pathogen's cell envelope.
Schnappinger
Dirk
D
Department of Microbiology and Immunology, Cornell University, 1300 York Avenue, New York, NY 10021, USA. dis2003@med.cornell.edu
Ehrt
Sabine
S
Voskuil
Martin I
MI
Liu
Yang
Y
Mangan
Joseph A
JA
Monahan
Irene M
IM
Dolganov
Gregory
G
Efron
Brad
B
Butcher
Philip D
PD
Nathan
Carl
C
Schoolnik
Gary K
GK
D000818:Animals; D015964:Gene Expression Regulation, Bacterial; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000378:metabolism*; D010588:Phagosomes / Q000382:microbiology*; D012329:RNA, Bacterial / Q000235:genetics; D014158:Transcription, Genetic
D012329:RNA, Bacterial