|Margarita O. Shleeva
Professor, Dr.Sci. (Biology)
Head of Laboratory
INBI, build. 1
+7 (495) 954-40-47
Directions of the investigations:
- Study of biochemical mechanisms of bacteria adaptation to stressfull conditions
- Study of biochemistry and physiology of dormant bacteria (mycobacteria)
- Development of experimental models of latent tuberculosis in vitro
- Trnascriptomic and proteomic analysis of dormant mycobacteria
- Study of the mechanisms of transition of viable bacteria to the dormancy and resuscitation from it.
- Search for new substances active against dormant form of mycobacteria
- Development of new approaches to diagnostic of tuberculosis
Most significant results:
1. In the field of dormant mycobacteria
- we have found that several members of the Actinomycetales (Micrococcus luteus, Rhodococcus rhodochrous and Mycobacterium tuberculosis(MTB) and Mycobacterium smegmatis) can enter to a “non-culturable” , dormant state in vitro.
- a number of models for transition of viable bacteria ( including Mycobacterium tuberculosis) to dormant state has been established
- morphological and biochemical properties of mycobacteria in dormant state were described
- transcriptomic and proteomic analysis revealed cohort of transcripts and proteins which demonstrated stability during long period of dormancy
- participation of toxin-antitoxin modules in formation of mycobacterial dormancy was experimentally proved
2. In the field of resuscitation of dormant bacteria
- in collaboration with Wales University, Aberystwyth (UK) we discovered of a family of bacterial secreted proteins (Rpf- lytic for bacterial peptidoglycan enzymes ) involved in controlling exit of mycobacteria from dormancy.
- Rpf proteins revealed protection activity toward mice tuberculosis when used as subunit vaccines. Rpf proteins participate in formation in cellular immunity against tuberculosis.
- Rpf-like proteins of M.tuberculosis are essential in replication of MTB cells in vivo (mice model).
- Rpf-like proteins of M. tuberculosis are important for reactivation of chronic tuberculosis infection in vivo. This makes Rpf proteins to be a perspective target for development of new class of anti-TB compounds and construction of new live vaccines on the basis of attenuated strains of MTB with deleted rpf genes.
- large fragments of peptidoglycan and individual muropeptides promote resuscitation of dormant mycobacteria. Rip A – an endopeptidase acting in conjunction with Rpf plays an important role in such resuscitation. This makes possible indirect action of Rpf as a resuscitation promoting factor.
- free unsaturated fatty acids can trigger resuscitation of dormant mycobacteria through activation of an intracellular adenylyl cyclase.
3. In the field of antimicrobial compounds
- we found new compounds (ortothiocyanate nitroaryles) which inhibit both Rpf enzymatic activity and resuscitation of dormant mycobacteria in vitro and in vivo (latent mice infection)
- novel class of thyopiridines are found to be sterilizing for dormant M.tuberculosis
- novel protonated polydialylamines demonstrate high sterilizing activity against mycobacteria
- Mukamolova_GV, Kaprelyants_AS, Young_DI, Young_M, Kell_DB A bacterial cytokine. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95:.8916-8921
- Mukamolova GV., Turapov OA, Young D, Kaprelyants AS, Kell DB& Young M. A family of autocrine growth factors in Mycobacterium tuberculosis. MOLECULAR MICROBIOLOGY 2002, 46: 623-635.
- Shleeva M. O., Bagramyan К, Telkov M. V., Mukamolova G. V., Young М, Kell D. B.and Kaprelyants A. S. Formation and resuscitation of “non-culturable” cells of Rhodococcus rhodochrous and Mycobacterium tuberculosis in prolonged stationary phase MICROBILOGY , 2002, 148:1581-1591
- Mukamolova GV., Kaprelyants AS, Kell DB& Young M. Adoption of the transiently non-culturable state- a bacterial survival strategy? ADVANCES IN MICROB.PHYSIOL. 2003, 47, 65-129
- Shleeva M., Mukamolova G., Young М, H.Williams and Kaprelyants A. S. Formation of “non-culturable” cells od Mycobacterium smegmatis in stationary phase in response to growth under suboptimal conditions and their Rpf-mediated resuscitation. MICROBIOLOGY-UK, 2004, 150:1687-1697
- Kuznetsov B.A., Davydova M.E., Shleeva M.O., Kaprelyants A.S., Yaropolov A.I. Electrochemical investigation of the dynamics of Mycobacterium smegmatis cells’ transformation to dormant, nonculturable form. BIOELECTROCHEMISTRY, 2004, 64: 125-131.
- Young M, Mukamolova G.V. and Kaprelyants A.S. Mycobacterial dormancy and its relation to persistence. IN: MYCOBACTERIAL MOLECULAR MICROBIOLOGY , ed. T.Parish, Horizon Scientific Press Ltd, 2005
- Katrina Downing, Vladimir Mischenko, Margarita Shleeva, Danielle Young, Michael Young, Arseny Kaprelyants, Alexander Apt, and Мalerie Mizrahi. Mutants of Mycobacterium tuberculosis lacking three of the five rpf-like genes are defective for growth in vivo and for resuscitation in vitro. Infrction and Immunity. , 2005, 73, N5.
- Galina V. Mukamolova, Alexey G. Murzin, Elena G. Salina, Galina R. Demina, Douglas B. Kell, Arseny S. Kaprelyants, Michael Young. Muralytic activity of Micrococcus luteus Rpf and its relationship to physiological activity in promoting bacterial growth and resuscitation. MOLECULAR MICROBIOLOGY(2006) v.59, p.84-98.
- 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, 67, 672-684.
- 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, еВ174. 4.1
- 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.
- 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. 3.8
- 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. 3.1
- 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- 3.5
- 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 -3.5
- 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.- 1.9
- Nikitushkin VD, Demina GR, Shleeva MO, Kaprelyants AS. Peptidoglycan fragments stimulate resuscitation of “non-culturable” mycobacteria. ANTONIE VAN LEEUWENHOEK. 2013,103, 37-46
- Shleeva M., Goncharenko A., Kudykina Yu., Young D., Young M. and Kaprelyants A. Cyclic AMP-dependent Resuscitation of Dormant Mycobacteria by Exogenous Free Fatty Acids. PLOS ONE (2013), v.8, 12, e82914.
- Toxin-antitoxin vapBC locus participates in formation of the dormant state in Mycobacterium smegmatis FEMS Letters (2014), Volume: 352 Issue: 1 Pages: 69-77
- Salina EG, Waddell SJ, Hoffmann N, Rosenkrands I, Butcher PD, Kaprelyants AS. Potassium availability triggers Mycobacterium tuberculosis transition to, and resuscitation from, non-culturable (dormant) states. Open Biology 2014, 4(10): 140106
- Margarita Shleeva, Tatiana Kondratieva, Elvira Rubakova, Galina Vostroknutova, Arseny Kaprelyants and Alexander Apt. Reactivation of dormant “non-culturable” Mycobacterium tuberculosis developed in vitro after injection in mice: both the dormancy depth and host genetics influence the outcome. Microbial Pathogenesis.(2014) doi:10.1016/j.micpath.2014.11.016
- Shleeva M, Kondratieva T, Rubakova E, Vostroknutova G, Kaprelyants A, Apt A. Reactivation of dormant “non-culturable” Mycobacterium tuberculosis developed in vitro after injection in mice: both the dormancy depth and host genetics influence the outcome. Microb Pathog. 2015 ;78:63-6.
- Timofeeva LM, Kleshcheva NA, Shleeva MO, Filatova MP, Simonova YA, Ermakov YA, Kaprelyants AS. Nonquaternary poly(diallylammonium) polymers with different amine structure and their biocidal effect on Mycobacterium tuberculosis and Mycobacterium smegmatis. Appl Microbiol Biotechnol. 2015; 99(6):2557-71.
- Nikitushkin VD, Demina GR, Shleeva MO, Guryanova SV, Ruggiero A, Berisio R, Kaprelyants AS. A product of RpfB and RipA joint enzymatic action promotes the resuscitation of dormant mycobacteria. FEBS J. 2015; 282(13):2500-11.
- Dmitriy V. Ignatov, Elena G. Salina, Mikhail V. Fursov, Timofey A. Skvortsov, Tatyana L. Azhikina, Arseny S. Kaprelyants (2015) Dormant non-culturable Mycobacterium tuberculosis retains stable low-abundant mRNA. BMC Genomics, 16:954