Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

  • Print ISSN 0367-6722

  • Online ISSN 0976-0555

  • NAAS Rating 6.43

  • SJR 0.263

  • Impact Factor 0.5 (2023)

Frequency :
Monthly (January, February, March, April, May, June, July, August, September, October, November and December)
Indexing Services :
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical Abstracts, Indian Science Abstracts, EBSCO Indexing Services, Index Copernicus
Submit

Research Article

Indian Journal of Animal Research, volume 55 issue 7 (july 2021) : 774-779

Regulatory Role of fnr Gene in Growth and tolA Gene Expression in Salmonella Typhimurium

Nikhil K.C., Swagatika Priyadarsini, M. Pashupathi, Barkha Ratta, Meeta Saxena, S. Ramakrishnan, Parthasarathi Behera, Ajay Kumar
1Division of Animal Biochemistry, IVRI, Izatnagar-243 122, Bareilly, Uttar Pradesh, India.
Cite article:- K.C. Nikhil, Priyadarsini Swagatika, Pashupathi M., Ratta Barkha, Saxena Meeta, Ramakrishnan S., Behera Parthasarathi, Kumar Ajay (2021). Regulatory Role of fnr Gene in Growth and tolA Gene Expression in Salmonella Typhimurium. Indian Journal of Animal Research. 55(7): 774-779. doi: 10.18805/IJAR.B-4120.

ABSTRACT

Background: Salmonella Typhimurium (S.Typhimurium) adapts to the broad fluctuations of oxygen concentrations encountered in the host. The transition from aerobic to microaerobic/anaerobic condition encountered in the intestine is mainly regulated by fumarate and nitrate reductase (fnr) regulatory gene and aerobic respiratory control A (arcA) gene. Aim is to appraise the role of fnr gene under anaerobic conditions.
Methods: In this study, we deleted fnr gene from S.Typhimurium using lambda red-recombinase mediated gene knockout protocol. Further carried out in vitro characterization and analyzed the differential protein expression in wild type (WT) and isogenic Δfnr null mutant (Δfnr) using SDS-PAGE and MALDI-TOF mass spectrometry under anaerobic conditions.
Result: In growth competition, WT strain outcompeted the Δfnr and biofilm-forming ability of Δfnr was significantly reduced compared to WT strain. Swimming motility was reduced in Δfnr strain. Besides, differential protein expression revealed the global changes in the expression of many proteins in fnr strain. One differentially expressed protein was identified as TolA, an inner membrane envelope protein. It points out that fnr may regulate the genes responsible for motility and biofilm formation. FNR protein positively regulates TolA, which is important for bacterial virulence, maintenance of membrane integrity, LPS production and replication of bacteria.

REFERENCES

  1. Behera, P., Nikhil, K.C., Kumar, A., Gali, J.M., De, A., Mohanty, A.K., Ali, M.A. and Sharma, B. (2020). Comparative proteomic analysis of Salmonella Typhimurium wild type and its isogenic fnr null mutant during anaerobiosis reveals new insight into bacterial metabolism and virulence. Microb. Pathog. 140: 103936. 
  2. Chakraborty, S., Roychoudhury, P., Samanta, I., Subudhi, P.K., Das, M., De, A., Bandyopadhayay, S., Joardar, S.N., Mandal, M. and Qureshi, A. (2020). Molecular detection of biofilm, virulence and antimicrobial resistance associated genes of Salmonella serovars isolated from pig and chicken of Mizoram, India. Indian J. Anim. Res. 54(5).
  3. Chakroun, I., Cordero, H., Mahdhi, A., Morcillo, P., Fedhila, K., Cuesta, A., Bakhrouf, A., Mahdouani, K. and Esteban, M.Á. (2017). Adhesion, invasion, cytotoxic effect and cytokine production in response to atypical Salmonella Typhimurium infection. Microb. Pathog. 106: 40-49.
  4. Chakroun, I., Mahdhi, A., Morcillo, P., Cordero, H., Cuesta, A., Bakhrouf, A., Mahdouani, K. and Esteban, M.Á. (2018). Motility, biofilm formation, apoptotic effect and virulence gene expression of atypical Salmonella Typhimurium outside and inside Caco-2 cells. Microb. Pathog. 114: 153-162.
  5. Chelvam, K.K., Chai, L.C. and Thong, K.L. (2014). Variations in motility and biofilm formation of Salmonella enterica serovar Typhi. Gut Pathog. 6(1): 2.
  6. Chopra, A.K., Peterson, J.W., Chary, P. and Prasad, R. (1994). Molecular characterization of an enterotoxin from Salmonella typhimurium. Microb. Pathog. 16(2): 85-98.
  7. Crack, J., Green, J. and Thomson, A.J. (2004). Mechanism of oxygen sensing by the bacterial transcription factor fumarate-nitrate reduction (FNR). J. Biol. Chem. 279(10): 9278-9286.
  8. Datsenko, K.A. and Wanner, B.L. (2000). One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl. Acad. Sci. 97(12): 6640-6645.
  9. Fernández, P., Velásquez, F., Garcias-Papayani, H., Amaya, F.A., Ortega, J., Gómez, S., Santiviago, C.A. and Álvarez, S.A. (2018). Fnr and ArcA regulate lipid a hydroxylation in Salmonella enteritidis by controlling lpxO expression in response to oxygen availability. Front. Microbiol. 9: 1220.
  10. Fink, R.C., Evans, M.R., Porwollik, S., Vazquez-Torres, A., Jones-Carson, J., Troxell, B., Libby, S.J., McClelland, M. and Hassan, H.M. (2007). FNR is a global regulator of virulence and anaerobic metabolism in Salmonella enterica serovar Typhimurium (ATCC 14028s). J. Bacteriol. 189(6): 2262-2273.
  11. Finlay, B.B. and Falkow, S. (1990). Salmonella interactions with polarized human intestinal Caco-2 epithelial cells. J. Infect. Dis. 162(5): 1096-1106.
  12. Grantcharova, N., Peters, V., Monteiro, C., Zakikhany, K. and Römling, U. (2010). Bistable expression of CsgD in biofilm development of Salmonella enterica serovar typhimurium. J. Bacteriol. 192(2): 456-466.
  13. Gunsalus, R.P. and Park, S.-J. (1994). Aerobic-anaerobic gene regulation in Escherichia coli: control by the ArcAB and Fnr regulons. Res. Microbiol. 145(5-6): 437-450.
  14. Gupta, A.R., Dey, S., Saini, M. and Swarup, D. 2014. Down-regulation of expression of type 1 collagen gene in dentin of fluoride exposed rats. Indian J. Anim. Res. 48(6): 601-604.
  15. HARRIS, J.C., DUPONT, H.L. and HORNICK, R.B. (1972). Fecal leukocytes in diarrheal illness. Ann. Intern. Med. 76(5): 697-703.
  16. Hassan, H.M. and Sun, H.C. (1992). Regulatory roles of Fnr, Fur and Arc in expression of manganese-containing superoxide dismutase in Escherichia coli. Proc. Natl. Acad. Sci. 89(8): 3217-3221.
  17. Iyoda, S., Kamidoi, T., Hirose, K., Kutsukake, K. and Watanabe, H. (2001). A flagellar gene fliZ regulates the expression of invasion genes and virulence phenotype in Salmonella enterica serovar Typhimurium. Microb. Pathog. 30(2): 81-90.
  18. Khoramian-Falsafi, T., Harayama, S., Kutsukake, K. and Pechere, J.C. (1990). Effect of motility and chemotaxis on the invasion of Salmonella typhimurium into HeLa cells. Microb. Pathog. 9(1): 47-53.
  19. Khoroshilova, N., Popescu, C., Münck, E., Beinert, H. and Kiley, P.J. (1997). Iron-sulfur cluster disassembly in the FNR protein of Escherichia coli by O2:[4Fe-4S] to [2Fe-2S] conversion with loss of biological activity. Proc. Natl. Acad. Sci. 94(12): 6087-6092.
  20. Lee, C.A. and Falkow, S. (1990). The ability of Salmonella to enter mammalian cells is affected by bacterial growth state. Proc. Natl. Acad. Sci. 87(11): 4304-4308.
  21. Lombardo, M.-J., Bagga, D. and Miller, C.G. (1991). Mutations in rpoA affect expression of anaerobically regulated genes in Salmonella typhimurium. J. Bacteriol. 173(23): 7511-7518.
  22. Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265-275.
  23. Masilamani, R., Cian, M.B. and Dalebroux, Z.D. (2018). Salmonella Tol-Pal reduces outer membrane glycerophospholipid levels for envelope homeostasis and survival during bacteremia. Infect. Immun. 86(7): e00173-18.
  24. Miki, T., Okada, N., Shimada, Y. and Danbara, H. (2004). Characterization of Salmonella pathogenicity island 1 type III secretion-dependent hemolytic activity in Salmonella enterica serovar Typhimurium. Microb. Pathog. 37(2): 65-72.
  25. Monteiro, C., Papenfort, K., Hentrich, K., Ahmad, I., Le Guyon, S., Reimann, R., Grantcharova, N. and Römling, U. (2012). Hfq and Hfq-dependent small RNAs are major contributors to multicellular development in Salmonella enterica serovar Typhimurium. RNA Biol. 9(4): 489-502.
  26. Morimoto, Y. V, Namba, K. and Minamino, T. (2017). Measurements of free-swimming speed of motile Salmonella cells in liquid media. Bio-protocol 7: e2093.
  27. Nair, A., Rawool, D.B., Doijad, S., Poharkar, K., Mohan, V., Barbuddhe, S.B., Kolhe, R., Kurkure, N. V, Kumar, A. and Malik, S.V.S. (2015). Biofilm formation and genetic diversity of Salmonella isolates recovered from clinical, food, poultry and environmental sources. Infect. Genet. Evol. 36: 424-433.
  28. Park, H.J., Kim, H.J., Park, S.H., Shin, E.G., Kim, J.H. and Kim, H.Y. (2008). Direct and quantitative analysis of Salmonella enterica serovar Typhimurium using real-time PCR from artificially contaminated chicken meat. J. Microbiol .Biotechnol. 18(8): 1453-1458.
  29. Park, S.J. and Gunsalus, R.P. (1995). Oxygen, iron, carbon and superoxide control of the fumarase fumA and fumC genes of Escherichia coli: role of the arcA, fnr and soxR gene products. J. Bacteriol. 177(21): 6255-6262.
  30. Paterson, G.K., Northen, H., Cone, D.B., Willers, C., Peters, S.E. and Maskell, D.J. (2009). Deletion of tolA in Salmonella Typhimurium generates an attenuated strain with vaccine potential. Microbiology 155(1): 220-228.
  31. Pegues, D.A. (2005). Salmonella species, including Salmonella typhi. Princ. Pract. Infect. Dis. 2636-2654.
  32. Sambrook, J. (2001). Molecular cloning: a laboratory manual/Joseph Sambrook, David W. Russell.
  33. Samhita, L., Nanjundiah, V. and Varshney, U. (2014). How many initiator tRNA genes does Escherichia coli need? J. Bacteriol. 196(14): 2607-2615.
  34. Shukla, S.K. and Rao, T.S. (2017). An Improved Crystal Violet Assay for Biofilm Quantification in 96-Well Microtitre Plate. bioRxiv 100214.
  35. Spiro, S. and Guest, J.R. (1990). FNR and its role in oxygen-regulated gene expression in Escherichia coli. FEMS Microbiol. Rev. 6(4): 399-428.
  36. Wing, H.J., Williams, S.M. and Busby, S.J. (1995). Spacing requirements for transcription activation by Escherichia coli FNR protein. J. Bacteriol. 177(23): 6704-6710.
In this Article
    Contact us
    Follow us

    Editorial Board

    View all (0)