THE ROLE OF mutR GENE IN METABOLISM AND VIRULENCE OF emm12 GENOTYPE STREPTOCOCCUS PYOGENES STRAINS

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  • Authors: Zutkis A.A.1, Milman B.L.1, Dmitriev A.V.2
  • Affiliations:
    1. Institute of Experimental Medicine of the North-West Branch of the Russian Academy of Medical Sciences, St. Petersburg, Russian Federation
    2. Institute of Experimental Medicine of the North-West Branch of the Russian Academy of Medical Sciences, St. Petersburg, Russian Federation 197376, Russian Federation, St. Petersburg, Аkademika Pavlova str., 12, Institute of Experimental Medicine. Phone: +7 (812) 234-68-57. Fax: +7 (812) 234-94-77
  • Issue: Vol 4, No 4 (2014)
  • Pages: 339-346
  • Section: ORIGINAL ARTICLES
  • URL: https://iimmun.ru/iimm/article/view/244
  • DOI: https://doi.org/10.15789/2220-7619-2014-4-339-346

Cite item

Abstract

In the present study the functional role of the mutR regulatory protein gene of Streptococcus pyogenes (emm12) was studied. The mutR gene was inactivated in the strains no. 97 and no. 152 by insertional mutagenesis. Inactivation of the mutR gene was found to affect the dynamic and characteristics of bacterial growth in liquid medium. Expression of secreted nucleases was significantly lower in the mutant strains compared to the wild-type strains. Two-dimensional electrophoresis and mass-spectrometry revealed differences in expression of number of the proteins in mutant strains
compared to the wild-type strains. Inactivation of the mutR gene negatively affected capacity of S. pyogenes to adhere to human epithelial cells. Finally, the virulence properties of the no. 152[mutR] mutant strains were found to be 4,7-fold less compared to the strain no. 152, while the no. 97[mutR] mutant strain became avirulent compared to the strain no. 97 due to insertional inactivation of the mutR gene.

About the authors

A. A. Zutkis

Institute of Experimental Medicine of the North-West Branch of the Russian Academy of Medical Sciences, St. Petersburg, Russian Federation

Email: fake@neicon.ru

Junior Researcher, Laboratory of the Functional Genomics and Proteomics of Microorganisms, Department of Molecular Microbiology, Institute of Experimental Medicine, St. Petersburg, Russian Federation

Russian Federation

B. L. Milman

Institute of Experimental Medicine of the North-West Branch of the Russian Academy of Medical Sciences, St. Petersburg, Russian Federation

Email: fake@neicon.ru

PhD, MD (Chemistry), Head of the Laboratory of Biomedical and Pharmaceutical Mass-Spectrometry, Department of Molecular Microbiology, Institute of Experimental Medicine, St. Petersburg, Russian Federation

Russian Federation

A. V. Dmitriev

Institute of Experimental Medicine of the North-West Branch of the Russian Academy of Medical Sciences, St. Petersburg, Russian Federation 197376, Russian Federation, St. Petersburg, Аkademika Pavlova str., 12, Institute of Experimental Medicine. Phone: +7 (812) 234-68-57. Fax: +7 (812) 234-94-77

Author for correspondence.
Email: admitriev10@yandex.ru

PhD, MD (Biology), Deputy Director on Science, Head of the Department of Ecological Physiology, Institute of Experimental Medicine, St. Petersburg, Russian Federation

Russian Federation

References

  1. Дмитриев А.В., Рождественская А.С., Зуткис А.А., Тотолян А.А. Направленная регуляция п 1. атогенных свойств стрептококков // Медицинский академ. журн. 2009. № 4. С. 50–58. [Dmitriev A.V., Rozhdestvenskaya A.S., Zutkis A.A., Totolian A.A. Targeted regulation of pathogenic properties in streptococci. Meditsinskii akademicheskii zhurnal = Medical Aсademic Journal, 2009, no. 4, pp. 50–58. (In Russ.)]
  2. Маниатис Т., Фрич Э., Сэмбрук Дж. Методы генетической инженерии. Молекулярное клонирование / Под ред. А.А. Баева, К.Г. Скрябина. М.: Мир, 1984. 479 с. [Maniatis T., Fritsch E.E., Sambrook J. Molecular cloning: a laboratory manual. N.Y.: Cold Spring Harbor Lab., 1982, 545 p.]
  3. Рождественская А.С., Дмитриев А.В., Грабовская К.Б., Тотолян А.А. Инактивация гена регулятора транскрипции Rgg изменяет экспрессию секретируемых факторов патогенности и вирулентность Streptococcus pyogenes // Медицинский академ. журн. 2008. № 2. С. 21–27. [Rozhdestvenskaja A.S., Dmitriyev A.V., Grabovskaja K.B., Totolian A.A. Inactivation of the transcription regulator gene Rgg leads to changes in the expression of secreted pathogenicity factors and the virulence of Streptococcus pyogenes. Meditsinskii akademicheskii zhurnal = Medical Aсademic Journal, 2008, no. 2, pp. 21–27. (In Russ.)]
  4. Рождественская А.С., Сантос-Санчес И., Дмитриев А.В. Природная мутация в гене белка-регулятора BgrR, приводящая к нарушению синтеза белка Bac, фактора патогенности Streptococcus agalactiae // Инфекция и иммунитет. 2013. № 1. С. 43–48. [Rozhdestvenskaya A.S., Santos-Sanches I., Dmitriev A.V. Natural mutation in the gene of response regulator BgrR resulting in repression of Bac protein synthesis, a pathogenicity factor of Streptococcus agalactiae. Infektsiya i immunitet = Russian Journal of Infection and Immunity, 2013, no. 1, pp. 43–48. (In Russ.)]
  5. Anbalagan S., McShan W.M., Dunman P.M., Chaussee M.S. Identification of Rgg binding sites in the Streptococcus pyogenes chromosome. J. Bacteriol., 2011, vol. 193, no. 18, pp. 4933–4942.
  6. Beckert S., Kreikemeyer B., Podbielski A. Group A streptococcal rofA gene is involved in the control of several virulence genes and eukaryotic cell attachment and internalization. Infect. Immun., 2001, vol. 69, no. 1, pp. 534–537.
  7. Chang J.C., LaSarre B., Jimenez J.C., Aggarwal C., Federle M.J. Two group A streptococcal peptide pheromones act through opposing Rgg regulators to control biofilm development. PLoS Pathog., 2011, vol. 7, no. 8.
  8. Chaussee M.A., Callegari E.A., Chaussee M.S. Rgg regulates growth phase-dependent expression of proteins associated with secondary metabolism and stress in Streptococcus pyogenes. J. Bacteriol., 2004, vol. 186, no. 21, pp. 7091–7099.
  9. Chaussee M.S., Somerville G.A., Reitzer L., Musser J.M. Rgg coordinates virulence factor synthesis and metabolism in Streptococcus pyogenes. J. Bacteriol., 2003, vol. 185, no. 20, pp. 6016–6024.
  10. Connolly K.L., Braden A.K., Holder R.C., Reid S.D. Srv mediated dispersal of streptococcal biofilms through SpeB is observed in CovRS+ strains. PLoS One, 2011, vol. 6, no. 12, p. e28640.
  11. Cunningham M.W. Pathogenesis of group A streptococcal infections. Clin. Microbiol. Rev., 2000, vol. 13, no. 3, pp. 470–511.
  12. Cusumano Z.T., Watson M.E., Caparon M.G. Streptococcus pyogenes arginine and citrulline catabolism promotes infection and modulates innate immunity. Infect. Immun., 2014, vol. 82, no. 1, pp. 233–242.
  13. Dmitriev A.V., McDowell E.J., Chaussee M.S. Inter- and intraserotypic variation in the Streptococcus pyogenes Rgg regulon. FEMS Microbiol. Lett., 2008, vol. 284, no. 1, pp. 43–51.
  14. Dmitriev A.V., McDowell E.J., Kappeler K.V., Chaussee M.A., Chaussee M.S. The Rgg regulator of Streptococcus pyogenes influences utilization of nonglucose carbohydrates, prophage induction, and expression of the NAD-glycohydrolase virulence operon. J. Bacteriol., 2006, vol. 188, no. 20, pp. 7230–7241.
  15. Hause L.L., McIver K.S. Nucleotides critical for the interaction of the Streptococcus pyogenes Mga virulence regulator with Mgaregulated promoter sequences. J. Bacteriol., 2012, vol. 194, no. 18, pp. 4904–4919.
  16. Hondorp E.R., Hou S.C., Hempstead A.D., Hause L.L., Beckett D.M., McIver K.S. Characterization of the group A Streptococcus Mga virulence regulator reveals a role for the C-terminal region in oligomerization and transcriptional activation. Mol. Microbiol., 2012, vol. 83, no. 5, pp. 953–967.
  17. Kreikemeyer B., McIver K.S., Podbielski A. Virulence factor regulation and regulatory networks in Streptococcus pyogenes and their impact on pathogen-host interactions. Trends Microbiol., 2003, vol. 11, no. 5, pp. 224–232.
  18. Kwinn L.A., Khosravi A., Aziz R.K., Timmer A.M., Doran K.S., Kotb M., Nizet V. Genetic characterization and virulence role of the RALP3/LSA locus upstream of the streptolysin s operon in invasive M1T1 group A Streptococcus. J. Bacteriol., 2007, vol. 189, no. 4, pp. 1322–1329.
  19. Pulliainen A.T., Hytönen J., Haataja S., Finne J. Deficiency of the Rgg regulator promotes H2O2 resistance, AhpCF-mediated H2O2 decomposition, and virulence in Streptococcus pyogenes. J. Bacteriol., 2008, vol. 190, no. 9, pp. 3225–3235.
  20. Ribardo D.A., McIver K.S. Defining the Mga regulon: Comparative transcriptome analysis reveals both direct and indirect regulation by Mga in the group A streptococcus. Mol. Microbiol., 2006, vol. 62, no. 2, pp. 491–508.
  21. Roberts S.A., Scott J.R. RivR and the small RNA RivX: the missing links between the CovR regulatory cascade and the Mga regulon. Mol. Microbiol., 2007, vol. 66, no. 6, pp. 1506–1522.
  22. Siemens N., Fiedler T., Normann J., Klein J., Münch R., Patenge N., Kreikemeyer B. Effects of the ERES pathogenicity region regulator Ralp3 on Streptococcus pyogenes serotype M49 virulence factor expression. J. Bacteriol., 2012, vol. 194, no. 14, pp. 3618–3626.
  23. Siemens N., Kreikemeyer B. Heterologous expression of Ralp3 in Streptococcus pyogenes M2 and M6 strains affects the virulence characteristics. PLoS One, 2013, vol. 8, no. 2.
  24. Toukoki C., Gold K.M., McIver K.S., Eichenbaum Z. MtsR is a dual regulator that controls virulence genes and metabolic functions in addition to metal homeostasis in the group A streptococcus. Mol. Microbiol., 2010, vol. 76, no. 4, pp. 971–989.
  25. Treviño J., Liu Z., Cao T.N., Ramirez-Peña E., Sumby P. RivR is a negative regulator of virulence factor expression in group A Streptococcus. Infect. Immun., 2013, vol. 81, no. 1, pp. 364–372.
  26. Voyich J.M., Sturdevant D.E., Braughton K.R., Kobayashi S.D., Lei B., Virtaneva K., Dorward D.W., Musser J.M., DeLeo F.R. Genome-wide protective response used by group A Streptococcus to evade destruction by human polymorphonuclear leukocytes. Proc. Natl. Acad. Sci. USA, 2003, vol. 100, no. 4, pp. 1996–2001.
  27. Zutkis A.A., Dmitriev A.V., Chaussee M.S., Totolian A.A. Inactivation of the transcriptional regulator mutR gene affects virulent phenotype of Streptococcus pyogenes. Clin. Microbiol. Infect., 2009, vol. 15, no. 4, p. 1840.

Copyright (c) 2015 Zutkis A.A., Milman B.L., Dmitriev A.V.

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