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Phenotypic characterization of a conserved inner membrane protein YhcB in Escherichia coli

  • Microbial Physiology and Biochemistry
  • Published:
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Abstract

Although bacteria have diverse membrane proteins, the function of many of them remains unknown or uncertain even in Escherichia coli. In this study, to investigate the function of hypothetical membrane proteins, genome-wide analysis of phenotypes of hypothetical membrane proteins was performed under various envelope stresses. Several genes responsible for adaptation to envelope stresses were identified. Among them, deletion of YhcB, a conserved inner membrane protein of unknown function, caused high sensitivities to various envelope stresses and increased membrane permeability, and caused growth defect under normal growth conditions. Furthermore, yhcB deletion resulted in morphological aberration, such as branched shape, and cell division defects, such as filamentous growth and the generation of chromosome-less cells. The analysis of antibiotic susceptibility showed that the yhcB mutant was highly susceptible to various anti-folate antibiotics. Notably, all phenotypes of the yhcB mutant were completely or significantly restored by YhcB without the transmembrane domain, indicating that the localization of YhcB on the inner membrane is dispensable for its function. Taken together, our results demonstrate that YhcB is involved in cell morphology and cell division in a membrane localization-independent manner.

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References

  • Akerlund, T., Bernander, R., and Nordström, K. 1992. Cell division in Escherichia coli minB mutants. Mol. Microbiol.6, 2073–2083.

    Article  CAS  PubMed  Google Scholar 

  • Akerlund, T., Nordström, K., and Bernander, R. 1993. Branched Escherichia coli cells. Mol. Microbiol.10, 849–858.

    Article  CAS  PubMed  Google Scholar 

  • Choi, J.S., Kim, W., Suk, S., Park, H., Bak, G., Yoon, J., and Lee, Y. 2018. The small RNA, SdsR, acts as a novel type of toxin in Escherichia coli. RNA Biol.15, 1319–1335.

    Article  PubMed  PubMed Central  Google Scholar 

  • Choi, U. and Lee, C.R. 2019. Distinct roles of outer membrane porins in antibiotic resistance and membrane integrity in Escherichia coli. Front. Microbiol.10, 953.

    Article  PubMed  PubMed Central  Google Scholar 

  • Choi, U., Park, Y.H., Kim, Y.R., Seok, Y.J., and Lee, C.R. 2017. Effect of the RNA pyrophosphohydrolase RppH on envelope integrity in Escherichia coli. FEMS Microbiol. Lett.364, fnx152.

    Article  Google Scholar 

  • 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. USA97, 6640–6645.

    Article  CAS  PubMed  Google Scholar 

  • Davie, E., Sydnor, K., and Rothfield, L.I. 1984. Genetic basis of minicell formation in Escherichia coli K-12. J. Bacteriol.158, 1202–1203.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hiraga, S., Niki, H., Ogura, T., Ichinose, C., Mori, H., Ezaki, B., and Jaffé, A. 1989. Chromosome partitioning in Escherichia coli: novel mutants producing anucleate cells. J. Bacteriol.171, 1496–1505.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Keseler, I.M., Mackie, A., Santos-Zavaleta, A., Billington, R., Bonavides-Martínez, C., Caspi, R., Fulcher, C., Gama-Castro, S., Kothari, A., Krummenacker, M., et al. 2017. The EcoCyc database: reflecting new knowledge about Escherichia coli K-12. Nucleic Acids Res.45, D543–D550.

    Article  CAS  PubMed  Google Scholar 

  • Lee, C.R., Kim, M., Park, Y.H., Kim, Y.R., and Seok, Y.J. 2014. RppH-dependent pyrophosphohydrolysis of mRNAs is regulated by direct interaction with DapF in Escherichia coli. Nucleic Acids Res.42, 12746–12757.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li, G., Hamamoto, K., and Kitakawa, M. 2012. Inner membrane protein YhcB interacts with RodZ involved in cell shape maintenance in Escherichia coli. ISRN Mol. Biol.2012, 304021.

    Article  PubMed  PubMed Central  Google Scholar 

  • Maddalo, G., Stenberg-Bruzell, F., Götzke, H., Toddo, S., Björkholm, P., Eriksson, H., Chovanec, P., Genevaux, P., Lehtiö, J., Ilag, L.L., et al. 2011. Systematic analysis of native membrane protein complexes in Escherichia coli. J. Proteome Res.10, 1848–1859.

    Article  CAS  PubMed  Google Scholar 

  • Mitchell, A.M., Srikumar, T., and Silhavy, T.J. 2018. Cyclic enterobacterial common antigen maintains the outer membrane permeability barrier of Escherichia coli in a manner controlled by YhdP. mBio9, e01321–18.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nelson, D.E., Ghosh, A.S., Paulson, A.L., and Young, K.D. 2002. Contribution of membrane-binding and enzymatic domains of penicillin binding protein 5 to maintenance of uniform cellular morphology of Escherichia coli. J. Bacteriol.184, 3630–3639.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nelson, D.E. and Young, K.D. 2000. Penicillin binding protein 5 affects cell diameter, contour, and morphology of Escherichia coli. J. Bacteriol.182, 1714–1721.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nelson, D.E. and Young, K.D. 2001. Contributions of PBP 5 and DD-carboxypeptidase penicillin binding proteins to maintenance of cell shape in Escherichia coli. J. Bacteriol.183, 3055–3064.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nichols, R.J., Sen, S., Choo, Y.J., Beltrao, P., Zietek, M., Chaba, R., Lee, S., Kazmierczak, K.M., Lee, K.J., Wong, A., et al. 2011. Phenotypic landscape of a bacterial cell. Cell144, 143–156.

    Article  CAS  Google Scholar 

  • Paradis-Bleau, C., Kritikos, G., Orlova, K., Typas, A., and Bernhardt, T.G. 2014. A genome-wide screen for bacterial envelope biogenesis mutants identifies a novel factor involved in cell wall precursor metabolism. PLoS Genet.10, e1004056.

    Article  PubMed  PubMed Central  Google Scholar 

  • Paradis-Bleau, C., Markovski, M., Uehara, T., Lupoli, T.J., Walker, S., Kahne, D.E., and Bernhardt, T.G. 2010. Lipoprotein cofactors located in the outer membrane activate bacterial cell wall polymerases. Cell143, 1110–1120.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Park, S., Yoon, J., Lee, C.R., Lee, J.Y., Kim, Y.R., Jang, K.S., Lee, K.H., and Seok, Y.J. 2019. Polar landmark protein HubP recruits flagella assembly protein FapA under glucose limitation in Vibrio vulnificus. Mol. Microbiol.112, 266–279.

    Article  CAS  PubMed  Google Scholar 

  • Popham, D.L. and Young, K.D. 2003. Role of penicillin-binding proteins in bacterial cell morphogenesis. Curr. Opin. Microbiol.6, 594–599.

    Article  CAS  PubMed  Google Scholar 

  • Potluri, L.P., de Pedro, M.A., and Young, K.D. 2012. Escherichia coli low-molecular-weight penicillin-binding proteins help orient septal FtsZ, and their absence leads to asymmetric cell division and branching. Mol. Microbiol.84, 203–224.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Spratt, B.G. 1975. Distinct penicillin binding proteins involved in the division, elongation, and shape of Escherichia coli K12. Proc. Natl. Acad. Sci. USA72, 2999–3003.

    Article  CAS  PubMed  Google Scholar 

  • Spratt, B.G. and Pardee, A.B. 1975. Penicillin-binding proteins and cell shape in E. coli. Nature254, 516–517.

    Article  CAS  PubMed  Google Scholar 

  • Trueba, F.J. and Woldringh, C.L. 1980. Changes in cell diameter during the division cycle of Escherichia coli. J. Bacteriol.142, 869–878.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wikler, M.A. and CLSI. 2018. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard. 11th edn. Clinical and Laboratory Standards Institute, Wayne, Pennsylvania, USA.

    Google Scholar 

  • Young, K.D. 2003. Bacterial shape. Mol. Microbiol.49, 571–580.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a research grant from the NRF funded by the Ministry of Science and ICT (number NRF-2018R1A1A1A05023049).

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  1. Department of Biological Sciences, Myongji University, Yongin, 17058, Republic of Korea

    Chul Gi Sung, Umji Choi & Chang-Ro Lee

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  1. Chul Gi Sung
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  2. Umji Choi
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  3. Chang-Ro Lee
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Correspondence to Chang-Ro Lee.

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Sung, C.G., Choi, U. & Lee, CR. Phenotypic characterization of a conserved inner membrane protein YhcB in Escherichia coli. J Microbiol. 58, 598–605 (2020). https://doi.org/10.1007/s12275-020-0078-4

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  • DOI: https://doi.org/10.1007/s12275-020-0078-4

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