LABORATORY OF BIOCHEMISTRY OF STRESSES IN MICROORGANISMS
Head of Laboratory:
professor, PhD, DrSci
Staff of Laboratory:
Ostrovsky D.N professor
Vostroknutova G.N., PhD
Gonachrenko A.V., PhD
Demina G.R., PhD
Shleeva M.O., PhD
Salina E.G.. PhD
Nikitushkin V., PhD
Ershov Yu. , PhD
Shumkov M., PhD
Fursov M., PhD student
The laboratory is involved in study of mechanisms of bacterial cell response to
different kind of stresses. In particular, we are interested in the mechanisms of cell survival under conditions of starvation and in stationary phase. The main attention is focused on formation of dormant and "non-culturable" cells of non-sporulating bacteria, their resuscitation and chemical and physical factors controlled these processes. Our current interest is associated with the study of dormant Mycobacteria tuberculosis (MTB) cells in connection with elucidation of mechanisms of latent tuberculosis. MTB, the causative agent of tuberculosis (TB), is responsible for more deaths than any other bacterial infectious disease. This organism is estimated to infect about one third of the Globe population. Nevertheless, ninety percent of these 2 billion people remain clinically healthy because persistent bacteria they harbor are quiescent, and these individuals never develop overt TB. Such latent infections are difficult to diagnose and treat. Latent infection in adults can reactivate at any time and is often accompanied by lung tissue destruction.
1. We have found that several members of the Actinomycetales (Micrococcus luteus, Rhodococcus rhodochrous and Mycobacterium tuberculosis and Mycobacterium smegmatis) can enter a "non-culturable" (NC) state when grown to and maintained in stationary phase in batch culture. These "non-culturable" organisms may be described as dormant, since they have extremely low metabolic activity and they require special treatment to promote their resuscitation.
2. In collaboration with Wales University, Aberystwyth (UK) we discovered a family of proteins involved in controlling dormancy and "non-culturability" in mycobacteria. Micrococcus luteus, a relative of M.tuberculosis, produces a "resuscitation-promoting factor" (Rpf) that is required to break the NC status of dormant organisms. Rpf homologues are widely distributed among the actinobacteria and multiple copies of rpf-like genes are found in M. tuberculosis as well as in other mycobacteria, corynebacteria and streptomycetes. Rpf resuscitates NC cells of M. tuberculosis as well as those of these other actinobacteria.
The structure of Rpf conserved domain shows strong resemblance to the c-type lysozyme fold. This suggests that the Rpf proteins have muralytic activity, what has been recently experimentally proved in this lab. The products of this activity are peptidoglican fragments liberated in the surrounding medium.
3. In experiments in vivo, we found that Rpf proteins control reactivation of latent TB infection in animals
4. Fragments of peptidoglican of mycobacterial cells stimulate reactivation of NC, dormant cells.
5. New inhibitors of Rpf resuscitation and enzymatic activities belonged to nitroaryles family have been found
6. Histon- like protein (HLP) participates in formation of dormant forms of M.smegmatis.
7. New anti-TB vaccine based on attenuated strains of MTB with knock-outed Rpfs has demonstrated high protective potential in animal experimental tuberculosis.
- capillary electrophoresis
- automatic registration of bacterial growth curves in 96-wells plates with Multiscan Accent
- flow cytometry
Institute of chemical physics, RAS
Institute of organic chemistry, RAS
Institute of microbiology, RAS
Institute of biochemistry and physiology of microorganisms, RAS
Central Institute of tuberculosis , RAMS
State Scientific Center for Applied Microbiology
Institute of Biological Sciences, University of Wales, Aberystwyth, UK
Manchester University (UMIST), UK
Centre of Excellence for Biomedical TB research, Johannesburg, South Africa
1. Nikitushkin VD, Demina GR, Shleeva MO, Kaprelyants AS.
Peptidoglycan fragments stimulate resuscitation of "non-culturable" mycobacteria. ANTONIE VAN LEEUWENHOEK. 2012 Aug 4. [Epub ahead of print]
2. Kaprelyants AS, Mukamolova GV, Ruggiero A, Makarov VA, Demina GR, Shleeva MO, Potapov VD, Shramko PA. Resuscitation-Promoting Factors (RPF): In Search of Inhibitors. PROTEIN PEPT LETT. 2012, 19(10), 1026-34.
3. Shleeva M. , Yulia K. Kudykina, Galina N. Vostroknutova, Natalia E. Suzina, Andrey L. Mulyukin, Arseny S. Kaprelyants. Dormant ovoid cells of Mycobacterium tuberculosis formed in response to gradual external acidification. TUBERCULOSIS, 2011, 91, 146-154
4. Kondratieva T., Rubakova E., Kana B., Biketov S., Potapov V., Kaprelyants A., Apt A.. Mycobacterium tuberculosis attenuated by multiple deletions of rpf genes effectively protects mice against TB infection. TUBERCULOSIS, 2011, 91, 219-223
5. Koltunov V, Greenblatt CL, Goncharenko AV, Demina GR, Klein BY, Young M, Kaprelyants AS. Structural Changes and Cellular Localization of Resuscitation-Promoting Factor in Environmental Isolates of Micrococcus luteus. MICROB. ECOL. 2010, 59, 296-310
6. Anuchin AM, Goncharenko AV, Demina GR, Mulyukin AL, Ostrovsky DN, Kaprelyants AS. The role of histone-like protein, Hlp, in Mycobacterium smegmatis dormancy. FEMS Microb.Letters, 2010, 308, 101-107.
7. Zhu K, Kaprelyants AS, Salina EG , Markx GH. Separation by dielectrophoresis of dormant and nondormant bacterial cells of Mycobacterium smegmatis. BIOMICROFLUIDICS, 2010, Jun 29; 4(2).
8. Young M., Artsatbanov V., Beller H.R., Chandra G., Chater K., Dover L., Goh E., Kahan T., Kaprelyants A., Kyrpides N., Mahillon J., Markowitz V., Mukamolova G., Oren A., Rokem S., Smith M., and Greenblatt C. Genome sequence of the Fleming strain of Micrococcus luteus, a simple free-living actinobacterium. J. BACTERIOLOGY, 2010, 192, 841-860.
9. Anuchin AM, Mulyukin AL, Suzina NE, Duda VI, El-Registan GI, and Kaprelyants AS. Dormant forms of Mycobacterium smegmatis with distinct morphology. MICROBIOLOGY-SGM (2009) 155, pp.1071-1079.
10. Demina G.R., Makarov V.A., Nikitushkin V.D., Ryabova O.B., Vostroknutova G.N., Salina E.G., Shleeva M.O., Goncharenko A.V., Kaprelyants A.S.. Finding of the Low Molecular Weight Inhibitors of Resuscitation Promoting Factor Enzymatic and Resuscitation Activity. (2009) PLOS ONE, 4, åÂ174.
11. Bavesh D. Kana, Bhavna G. Gordhan, Katrina J. Downing, Nackmoon Sung, Galina Vostroktunova, Edith E. Machowski, Liana Tsenova, Michael Young, Arseny Kaprelyants, Gilla Kaplan, Valerie Mizrahi. The resuscitation-promoting factors of Mycobacterium tuberculosis are required for virulence and resuscitation from dormancy but are collectively dispensable for growth in vitro. MOLECULAR MICROBIOLOGY, 2008, v.67, p.672-684
12. Mukamolova G., Salina E., Kaprelyants A. Mechanisms of latent tuberculosis: dormancy and resuscitation of Mycobacterium tuberculosis. , in NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES, NIH , Volume1. Frontiers in Research, HUMANA PRESS ,eds Vassil St. Georgiev, Karl Western, John McGowan. 2008, p.83-90.
|Last review: 11, March, 2013|
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