Cor interacts with outer membrane proteins to exclude FhuA-dependent phages
Superinfection exclusion (Sie) of FhuA-dependent phages is carried out by Cor in the Escherichia coli mEp167 prophage lysogenic strain. In this work, we present evidence that Cor is an outer membrane (OM) lipoprotein that requires the participation of additional outer membrane proteins (OMPs) to exclude FhuA-dependent phages. Two Cor species of ~13 and ~8.5 kDa, corresponding to the preprolipoprotein/prolipoprotein and lipoprotein, were observed by Western blot. Cell mutants for CorC17F, CorA18D and CorA57E lost the Sie phenotype for FhuA-dependent phages. A copurification affinity binding assay combined with LC_ESI_MS/MS showed that Cor bound to OMPs: OmpA, OmpC, OmpF, OmpW, LamB, and Slp. Interestingly, Sie for FhuA-dependent phages was reduced on Cor overexpressing FhuA+ mutant strains, where ompA, ompC, ompF, ompW, lamB, fhuE, genes were knocked out. The exclusion was restored when these strains were supplemented with plasmids expressing these genes. Sie was not lost in other Cor overexpressing FhuA+ null mutant strains JW3938(btuB-), JW5100(tolB-), JW3474(slp-). These results indicate that Cor interacts and requires some OMPs to exclude FhuA-dependent phages.
We especially thank the National Institute of Genetics (Japan) and Dr. Dimitris Georgellis for providing us BW25113 and the derivative knockout mutant strains (Keio Collection), and their corresponding plasmids (ASKA Collection). We also thank Dr. Thomas Silhavy for providing us Anti-LamB antibody. To Dr. Rosa Ma. Bermúdez Cruz, Dr. Jaime Ortega López, and two unknown reviewers for their critical observations and enriching discussions. To M. Sc. María Guadalupe Aguilar González for her support with nucleic acid sequencing and M. Sc. Marco A. Magos-Castro and M. Sc. Juan Carlos Osorio for their technical assistance. To Gabriela Mora for helping us in the administrative labor. This work was supported by Secretaria de Ciencia, Tecnología e Innovación de la Ciudad de México (SECYTI-CDMX), grant no. PICSA 11-107. E.S.A.H. was granted by CONACyT Mexico fellowship no. 210176.
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Conflict of interest
Authors: Emma S. Arguijo-Hernández, Javier Hernandez-Sanchez, Saida J. Briones-Peña, Norma Oviedo, Gabriel Guarneros and Luis Kameyama declare that have no conflicts of interest. Author: Guillermo Mendoza-Hernández passed away.
This article does not contain any studies with animals or human participants performed by any of the authors.
- 5.Reyes-Cortes R, Martinez-Penafiel E, Martinez-Perez F, de la Garza M, Kameyama L (2012) A novel strategy to isolate cell-envelope mutants resistant to phage infection: bacteriophage mEp213 requires lipopolysaccharides in addition to FhuA to enter Escherichia coli K-12. Microbiology 158:3063–3071CrossRefPubMedGoogle Scholar
- 7.Hernandez-Sanchez J, Bautista-Santos A, Fernandez L, Bermudez-Cruz RM, Uc-Mass A, Martinez-Penafiel E, Martinez MA, Garcia-Mena J, Guarneros G, Kameyama L (2008) Analysis of some phenotypic traits of feces-borne temperate lambdoid bacteriophages from different immunity groups: a high incidence of cor+, FhuA-dependent phages. Arch Virol 153:1271–1280CrossRefPubMedGoogle Scholar
- 15.Goldberg E, Grinius L, Letellier L (1994) Recognition, attachment, and injection. In: Karam JD (ed) Molecular biology of bacteriophage T4. American Society for Microbiology, Washington, DC, pp 347–356Google Scholar
- 45.Silhavy TJ, Berman, ML, Enquist LW (1984) Experiments with gene fusions. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
- 46.Polayes D, Hughes AJ (1994) Efficient protein expression and simple purification using the pPROEX-1 super(TM) system. FOCUS 16:81–84Google Scholar
- 47.Hanahan D (1985) Techniques for transformation of E. coli. In: Glover DM (ed) DNA cloning. IRL Press, Oxford, pp 109–135Google Scholar
- 58.Zhang H, Tang X, Munske GR, Zakharova N, Yang L, Zheng C, Wolff MA, Tolic N, Anderson GA, Shi L, Marshall MJ, Fredrickson JK, Bruce JE (2008) In vivo identification of the outer membrane protein OmcA-MtrC interaction network in Shewanella oneidensis MR-1 cells using novel hydrophobic chemical cross-linkers. J Proteome Res 7:1712–1720CrossRefPubMedPubMedCentralGoogle Scholar