Elements of signal transduction in Mycobacterium tuberculosis: In vitro phosphorylation in vivo expression of the response regulator MtrA

被引:94
作者
Via, LE [1 ]
Curcic, R [1 ]
Mudd, MH [1 ]
Dhandayuthapani, S [1 ]
Ulmer, RJ [1 ]
Deretic, V [1 ]
机构
[1] UNIV TEXAS, HLTH SCI CTR, DEPT MICROBIOL, SAN ANTONIO, TX 78284 USA
关键词
D O I
10.1128/jb.178.11.3314-3321.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A putative two-component system, mtrA-mtrB, was isolated from M. tuberculosis H37Rv by using phoB from Pseudomonas aeruginosa as a hybridization probe, The predicted gene product of mtrA displayed high similarity with typical response regulators, including AfsQ1, PhoB, PhoP, and OmpR. The predicted gene product of mtrB displayed similarities with the histidine protein kinases AfsQ2, PhoR, and EnvZ and other members of this class of proteins. Expression analysis in the T7 system showed that mtrA encoded a polypeptide with an apparent molecular mass of 30 kDa. MtrA was overproduced, purified, and demonstrated to participate in typical phosphotransfer reactions using heterologous histidine protein kinase, CheA, as a phosphoryl group donor, Mycobacterium bovis BCG, harboring an mtrA-gfp (green fluorescent protein cDNA) transcriptional fusion, was used to monitor mtrA expression iri infected J774 monolayers. Flow cytometric and fluorescence microscopic analyses indicated that the mtrA promoter was activated upon entry and incubation in J774 macrophages. In contrast, the hsp60-gp fusion displayed no change in expression under the growth conditions tested. These results suggest a potential role for mtrA in adaptation of the M. tuberculosis complex organisms to environmental changes which may include intracellular conditions.
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页码:3314 / 3321
页数:8
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