1887

Abstract

is a bacterium belonging to the class . It causes acute and chronic infections of the urogenital tract. The main features of this bacterium are an absence of cell wall and a reduced genome size (517–622 protein-encoding genes). Previously, we have isolated morphologically unknown colonies called micro-colonies (MCs) from the serum of patients with inflammatory urogenital tract infection.

MCs are functionally different from the typical colonies (TCs) in terms of metabolism and cell division.

To determine the physiological differences between MCs and TCs of and elucidate the pathways of formation and growth of MCs by a comparative proteomic analysis of these two morphological forms.

LC–MS proteomic analysis of TCs and MCs using an Ultimate 3000 RSLC nanoHPLC system connected to a QExactive Plus mass spectrometer.

The study of the proteomic profiles of colonies allowed us to reconstruct their energy metabolism pathways. In addition to the already known pentose phosphate and arginine deamination pathways, can utilise ribose phosphate and deoxyribose phosphate formed by nucleoside catabolism as energy sources. Comparative proteomic HPLC–MS analysis revealed that the proteomic profiles of TCs and MCs were different. We assume that MC cells preferably utilised deoxyribonucleosides, particularly thymidine, as an energy source rather than arginine or ribonucleosides. Utilisation of deoxyribonucleosides is less efficient as compared with that of ribonucleosides and arginine in terms of energy production. Thymidine phosphorylase DeoA is one of the key enzymes of deoxyribonucleosides utilisation. We obtained a DeoA overexpressing mutant that exhibited a phenotype similar to that of MCs, which confirmed our hypothesis.

In addition to the two known pathways for energy production (arginine deamination and the pentose phosphate pathway) can use deoxyribonucleosides and ribonucleosides. MC cells demonstrate a reorganisation of energy metabolism: unlike TC cells, they preferably utilise deoxyribonucleosides, particularly thymidine, as an energy source rather than arginine or ribonucleosides. Thus MC cells enter a state of energy starvation, which helps them to survive under stress, and in particular, to be resistant to antibiotics.

Funding
This study was supported by the:
  • Russian Science Foundation (Award 19-75-10124)
    • Principle Award Recipient: TatianaA. Semashko
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2022-01-17
2022-05-18
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References

  1. Waites K, Talkington D. New developments in human diseases due to mycoplasmas. In Blanchard A, Browning GF. eds Mycoplasmas Molecular 2005 pp 289–354
    [Google Scholar]
  2. Wong SS, Yuen KY. Acute pyelonephritis caused by Mycoplasma hominis . Pathology 1995; 27:61–63 [View Article] [PubMed]
    [Google Scholar]
  3. Rakovskaya IV, Gorina LG, Balabanov DN, Levina GA, Barhatova OI et al. Generalized Mycoplasma infection in patients and carriers. J Microbiol Immunol Infect 2013; 2:37–43
    [Google Scholar]
  4. Hasebe A, Mu HH, Cole BC. A potential pathogenic factor from Mycoplasma hominis is a TLR2-dependent, macrophage-activating, P50-related adhesin. Am J Reprod Immunol 2014; 72:285–295 [View Article] [PubMed]
    [Google Scholar]
  5. Baseman JB, Tully JG. Mycoplasmas: sophisticated, reemerging, and burdened by their notoriety. Emerg Infect Dis 1997; 3:21–32 [View Article]
    [Google Scholar]
  6. Levina GA, Barhatova OI, Gorina LG, Gamova NA, Goncharova SA et al. Unusual forms of Mycoplasma hominis persistence in the organism of infected people. J Microbiol Immunol Infect 2012; 4:104–109
    [Google Scholar]
  7. Rakovskaya IV, Ermolaeva SA, Levina GA, Barkhatova OI, Mukhachev AY et al. Microcolonies: a novel morphological form of pathogenic Mycoplasma spp. J Med Microbiol 2019; 68:1747–1758 [View Article] [PubMed]
    [Google Scholar]
  8. Ermolaeva SA, Rakovskaya IV, Miller GG, Sysolyatina EV, Mukhachev AY et al. Nonthermal plasma affects viability and morphology of Mycoplasma hominis and Acholeplasma laidlawii . J Appl Microbiol 2014; 116:1129–1136 [View Article]
    [Google Scholar]
  9. Gorbachev AY, Fisunov GY, Izraelson M, Evsyutina DV, Mazin PV et al. DNA repair in Mycoplasma gallisepticum . BMC Genomics 2013; 14:726 [View Article] [PubMed]
    [Google Scholar]
  10. Razin S, Rottem S. Techniques for the manipulation of Mycoplasma membranes . In Madd A. ed. Biochemical Analysis of Membranes Academic Press; 1976
    [Google Scholar]
  11. Senterfit L. Preparation of antigens and antisera. In Methods in Mycoplasmology vol. 2 1983 pp 401–404
    [Google Scholar]
  12. Waites KB, Bade DJ, Bébéar C, Brown SD, Davidson MK et al. Methods for antimicrobial susceptibility testing for human mycoplasmas; approved guideline. CLSI Publication Clinical and Laboratory Standards Institute 2011
    [Google Scholar]
  13. Mazin PV, Fisunov GY, Gorbachev AY, Kapitskaya KY, Altukhov IA et al. Transcriptome analysis reveals novel regulatory mechanisms in a genome-reduced bacterium. Nucleic Acids Res 2014; 42:13254–13268 [View Article] [PubMed]
    [Google Scholar]
  14. Semashko T, Arzamasov A, Evsyutina D, Fisunov G, Govorun V. Functions of restriction-modification system of Mycoplasma gallisepticum. Poster session presented at: World Microbe Forum; 2021 https://www.abstractsonline.com/cSubmit/submitSummary.asp?CKey=3A5E4F89-A519-46BC-9FFC-C87984CE2F8C&MKey=%7B10C6E7CD-1109-41C5-AA58-1C4EEA58AA09%7D
  15. Fisunov GY, Evsyutina DV, Semashko TA, Arzamasov AA, Manuvera VA et al. Binding site of MraZ transcription factor in Mollicutes. Biochimie 2016; 125:59–65 [View Article] [PubMed]
    [Google Scholar]
  16. Prjibelski A, Antipov D, Meleshko D, Lapidus A, Korobeynikov A. Using SPAdes De Novo Assembler. Curr Protoc Bioinformatics 2020; 70:e102 [View Article] [PubMed]
    [Google Scholar]
  17. Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics 2014; 30:2068–2069 [View Article] [PubMed]
    [Google Scholar]
  18. Shevchenko A, Wilm M, Vorm OM, Mann M. Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels. Anal Chem 1996; 68:850–858 [View Article] [PubMed]
    [Google Scholar]
  19. Zgoda V, Tikhonova O, Viglinskaya A, Serebriakova M, Lisitsa A et al. Proteomic profiles of induced hepatotoxicity at the subcellular level. Proteomics 2006; 6:4662–4670 [View Article] [PubMed]
    [Google Scholar]
  20. Ladefoged SA. Molecular dissection of Mycoplasma hominis . APMIS Suppl 2000; 97:1–45 [PubMed]
    [Google Scholar]
  21. Olson LD, Renshaw CA, Shane SW, Barile MF. Successive synovial Mycoplasma hominis isolates exhibit apparent antigenic variation. Infect Immun 1991; 59:3327–3329 [View Article] [PubMed]
    [Google Scholar]
  22. Christiansen G, Jensen LT, Boesen T, Emmersen J, Ladefoged SA et al. Molecular biology of Mycoplasma . Wien Klin Wochenschr 1997; 109:57–61
    [Google Scholar]
  23. Nyvold C, Birkelund S, Christiansen G. The Mycoplasma hominis P120 membrane protein contains a 216 amino acid hypervariable domain that is recognized by the human humoral immune response. Microbiology 1997; 143 (Pt 2):675–688 [View Article] [PubMed]
    [Google Scholar]
  24. Pollack JD. Carbohydrate metabolism and energy conservation. In Maniloff J. ed. Mycoplasmas: Molecular Biology and Pathogenesi Washington, DC, US: American Society for Microbiology; 1992 pp 181–200
    [Google Scholar]
  25. Pereyre S, Sirand-Pugnet P, Beven L, Charron A, Renaudin H et al. Life on arginine for Mycoplasma hominis: clues from its minimal genome and comparison with other human urogenital mycoplasmas. PLoS Genet 2009; 5:10 [View Article] [PubMed]
    [Google Scholar]
  26. Zelcer N, Huisman MT, Reid G, Wielinga P, Breedveld P et al. Evidence for two interacting ligand binding sites in human multidrug resistance protein 2 (ATP binding cassette C2). J Biol Chem 2003; 278:23538–23544 [View Article] [PubMed]
    [Google Scholar]
  27. Patzlaff JS, van der Heide T, Poolman B. The ATP/substrate stoichiometry of the ATP-binding cassette (ABC) transporter OpuA. J Biol Chem 2003; 278:29546–29551 [View Article] [PubMed]
    [Google Scholar]
  28. Davidson AL, Nikaido H. Overproduction, solubilization, and reconstitution of the maltose transport system from Escherichia coli . J Biol Chem 1990; 265:4254–4260 [PubMed]
    [Google Scholar]
  29. Sasaki Y, Ishikawa J, Yamashita A, Oshima K, Kenri T et al. The complete genomic sequence of Mycoplasma penetrans, an intracellular bacterial pathogen in humans. Nucleic Acids Res 2002; 30:5293–5300 [View Article] [PubMed]
    [Google Scholar]
  30. Sanders CC. Review of preclinical studies with ofloxacin. Clin Infect Dis 1992; 14:526–538 [View Article] [PubMed]
    [Google Scholar]
  31. Hahn FE, Sarre SG. Mechanism of action of gentamicin. J Infect Dis 1969; 119:364–369 [View Article]
    [Google Scholar]
  32. Piscitelli SC, Danziger LH, Rodvold KA. Clarithromycin and azithromycin: new macrolide antibiotics. Clin Pharm 1992; 11:137–152 [PubMed]
    [Google Scholar]
  33. Fenske JD, Kenny GE. Role of arginine deiminase in growth of Mycoplasma hominis . J Bacteriol 1976; 126:501–510 [View Article] [PubMed]
    [Google Scholar]
  34. Lewkowicz ES, Iribarren AM. Nucleoside phosphorylases. COC 2006; 10:1197–1215 [View Article]
    [Google Scholar]
  35. Hobby GL, Meyer K, Chaffee E. Observations on the mechanism of action of penicillin. Exp Biol Med 1942; 50:281–285 [View Article]
    [Google Scholar]
  36. Bigger JW. Treatment of staphylococcal infections with penicillin. Lancet 1944; 244:497–500
    [Google Scholar]
  37. Scherrer R, Moyed HS. Conditional impairment of cell division and altered lethality in hipA mutants of Escherichia coli K-12. J Bacteriol 1988; 170:3321–3326 [View Article] [PubMed]
    [Google Scholar]
  38. Shao Y, Harrison EM, Bi D, Tai C, He X et al. TADB: a web-based resource for Type 2 toxin-antitoxin loci in bacteria and archaea. Nucleic Acids Res 2011; 39:D606–11 [View Article] [PubMed]
    [Google Scholar]
  39. Yamaguchi Y, Park JH, Inouye M. Toxin–antitoxin systems in bacteria and archaea. Annu Rev Genet 2011; 45:61–79 [View Article]
    [Google Scholar]
  40. Amato SM, Orman MA, Brynildsen MP. Metabolic control of persister formation in Escherichia coli . Mol Cell 2013; 50:475–487 [View Article] [PubMed]
    [Google Scholar]
  41. Bernier SP, Lebeaux D, DeFrancesco AS, Valomon A, Soubigou G et al. Starvation, together with the SOS response, mediates high biofilm-specific tolerance to the fluoroquinolone ofloxacin. PLoS Genet 2013; 9:e1003144 [View Article] [PubMed]
    [Google Scholar]
  42. Fung DKC, Chan EWC, Chin ML, Chan RCY. Delineation of a bacterial starvation stress response network which can mediate antibiotic tolerance development. Antimicrob Agents Chemother 2010; 54:1082–1093 [View Article] [PubMed]
    [Google Scholar]
  43. Proctor RA, von Eiff C, Kahl BC, Becker K, McNamara P et al. Small colony variants: a pathogenic form of bacteria that facilitates persistent and recurrent infections. Nat Rev Microbiol 2006; 4:295–305 [View Article] [PubMed]
    [Google Scholar]
  44. Cottew GS. Pathogenicity of the subspecies mycoides of Mycoplasma mycoides for cattle, sheep and goats. Zentralbl Bakteriol Orig A 1979; 245:164–170 [PubMed]
    [Google Scholar]
  45. March JB, Clark J, Brodlie M. Characterization of strains of Mycoplasma mycoides subsp. mycoides small colony type isolated from recent outbreaks of contagious bovine pleuropneumonia in Botswana and Tanzania: evidence for a new biotype. J Clin Microbiol 2000; 38:1419–1425 [View Article] [PubMed]
    [Google Scholar]
  46. Pilo P, Frey J, Vilei EM. Molecular mechanisms of pathogenicity of Mycoplasma mycoides subsp. mycoides SC. Vet J 2007; 174:513–521 [View Article] [PubMed]
    [Google Scholar]
  47. Browning GF, Marenda MS, Noormohammadi AH, Markham PF. The central role of lipoproteins in the pathogenesis of mycoplasmoses. Vet Microbiol 2011; 153:44–50 [View Article] [PubMed]
    [Google Scholar]
  48. Jensen LT, Ladefoged S, Birkelund S, Christiansen G. Selection of Mycoplasma hominis PG21 deletion mutants by cultivation in the presence of monoclonal antibody 552. Infect Immun 1995; 63:3336–3347 [View Article] [PubMed]
    [Google Scholar]
  49. Ladefoged SA, Christiansen G. Mycoplasma hominis expresses two variants of a cell-surface protein, one a lipoprotein, and one not. Microbiology 1998; 144 (Pt 3):761–770 [View Article] [PubMed]
    [Google Scholar]
  50. Nyvold C, Birkelund S, Christiansen G. The Mycoplasma hominis P120 membrane protein contains a 216 amino acid hypervariable domain that is recognized by the human humoral immune response. Microbiology 1997; 143 (Pt 2):675–688 [View Article] [PubMed]
    [Google Scholar]
  51. Cacciotto C, Dessì D, Cubeddu T, Cocco AR, Pisano A et al. MHO_0730 as a surface-exposed calcium-dependent nuclease of Mycoplasma hominis promoting neutrophil extracellular trap formation and escape. J Infect Dis 2019; 220:1999–2008 [View Article] [PubMed]
    [Google Scholar]
  52. Goret J, Le Roy C, Touati A, Mesureur J, Renaudin H et al. Surface lipoproteome of Mycoplasma hominis PG21 and differential expression after contact with human dendritic cells. Future Microbiol 2016; 11:179–194 [View Article] [PubMed]
    [Google Scholar]
  53. Henrich B, Hopfe M, Kitzerow A, Hadding U. The adherence-associated lipoprotein P100, encoded by an opp operon structure, functions as the oligopeptide-binding domain OppA of a putative oligopeptide transport system in Mycoplasma hominis . J Bacteriol 1999; 181:4873–4878 [View Article] [PubMed]
    [Google Scholar]
  54. Hopfe M, Henrich B. OppA, the substrate-binding subunit of the oligopeptide permease, is the major ecto-ATPase of Mycoplasma hominis . J Bacteriol 2004; 186:1021–1928 [View Article] [PubMed]
    [Google Scholar]
  55. Hopfe M, Henrich B. OppA, the ecto-ATPase of Mycoplasma hominis induces ATP release and cell death in HeLa cells. BMC Microbiol 2008; 8:55 [View Article] [PubMed]
    [Google Scholar]
  56. Hopfe M, Dahlmanns T, Henrich B. In Mycoplasma hominis the OppA-mediated cytoadhesion depends on its ATPase activity. BMC Microbiol 2011; 11:185 [View Article] [PubMed]
    [Google Scholar]
  57. Citti C, Rosengarten R. Mycoplasma genetic variation and its implication for pathogenesis. Wien Klin Wochenschr 1997; 109:562–568 [PubMed]
    [Google Scholar]
  58. Chambaud I, Wróblewski H, Blanchard A. Interactions between mycoplasma lipoproteins and the host immune system. Trends in Microbiology 1999; 7:493–499 [View Article]
    [Google Scholar]
  59. Razin S, Yogev D, Naot Y. Molecular biology and pathogenicity of mycoplasmas. Microbiol Mol Biol Rev 1998; 62:1094–1156 [View Article] [PubMed]
    [Google Scholar]
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