期刊
ANTONIE VAN LEEUWENHOEK INTERNATIONAL JOURNAL OF GENERAL AND MOLECULAR MICROBIOLOGY
卷 103, 期 1, 页码 37-46出版社
SPRINGER
DOI: 10.1007/s10482-012-9784-1
关键词
Mycobacteria; Dormancy; Rpf; Reactivation; Peptidoglycan; Muropeptides
类别
资金
- Russian Academy of Sciences
- Federal Target Programme Molecular and cellular Biology Scientific and scientific-pedagogical personnel of innovative Russia [14740.1056, 14.740. 11.0246, 14.740.11.08.01]
- RFBR [11-04-00713-a, 11-04-01440-a]
Resuscitation promoting factors (Rpfs), belonging to a family of secreted actinobacterial proteins with predicted peptidoglycan (PG) hydrolytic activities, participate in the reactivation of dormant cells. In the present study we demonstrate that a recombinant truncated form of Micrococcus luteus Rpf hydrolyzes isolated PG of Mycobacterium smegmatis and Mycobacterium tuberculosis liberating PG fragments of different size. These fragments possess stimulatory activity toward non-culturable dormant M. smegmatis and M. tuberculosis cells, similar to the activity of recombinant Rpf. Relatively large PG fragments (0.1-0.5 mu m) obtained either by Rpf digestion or by PG ultrasonication revealed resuscitation activities when added in concentrations 0.1-0.2 mu g/ml to the resuscitation medium. It is suggested that PG fragments could either directly activate the resuscitation pathway of dormant mycobacteria or serve as a substrate for endogenous Rpf, resulting in low molecular weight products with resuscitation activity. Whilst both suggestions are plausible, it was observed that PG-dependent resuscitation activity was suppressed by means of a specific Rpf inhibitor (4-benzoyl-2-nitrophenylthiocyanate), which provides additional support for the second of these possibilities.
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