TolC-dependent secretion of an ankyrin repeat-containing protein of Rickettsia typhi

Simran J. Kaur, M. Sayeedur Rahman, Nicole Ammerman, Magda Beier-Sexton, Shane M. Ceraul, Joseph J. Gillespie, Abdu F. Azada

Research output: Contribution to journalArticle

Abstract

Rickettsia typhi, the causative agent of murine (endemic) typhus, is an obligate intracellular pathogen with a life cycle involving both vertebrate and invertebrate hosts. In this study, we characterized a gene (RT0218) encoding a C-terminal ankyrin repeat domain-containing protein, named Rickettsia ankyrin repeat protein 1 (RARP-1), and identified it as a secreted effector protein of R. typhi. RT0218 showed differential transcript abundance at various phases of R. typhi intracellular growth. RARP-1 was secreted by R. typhi into the host cytoplasm during in vitro infection of mammalian cells. Transcriptional analysis revealed that RT0218 was cotranscribed with adjacent genes RT0217 (hypothetical protein) and RT0216 (TolC) as a single polycistronic mRNA. Given one of its functions as a facilitator of extracellular protein secretion in some Gram-negative bacterial pathogens, we tested the possible role of TolC in the secretion of RARP-1. Using Escherichia coli C600 and an isogenic tolC insertion mutant as surrogate hosts, our data demonstrate that RARP-1 is secreted in a TolC-dependent manner. Deletion of either the N-terminal signal peptide or the C-terminal ankyrin repeats abolished RARP-1 secretion by wild-type E. coli. Importantly, expression of R. typhi tolC in the E. coli tolC mutant restored the secretion of RARP-1, suggesting that TolC has a role in RARP-1 translocation across the outer membrane. This work implies that the TolC component of the putative type 1 secretion system of R. typhi is involved in the secretion process of RARP-1.

Original languageEnglish (US)
Pages (from-to)4920-4932
Number of pages13
JournalJournal of Bacteriology
Volume194
Issue number18
DOIs
StatePublished - Sep 1 2012

Fingerprint

Rickettsia typhi
Ankyrin Repeat
Rickettsia
Proteins
Endemic Flea-Borne Typhus
Terminal Repeat Sequences
Escherichia coli
Secretory Pathway
Protein Transport
Invertebrates
Protein Sorting Signals
Life Cycle Stages
Genes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Kaur, S. J., Sayeedur Rahman, M., Ammerman, N., Beier-Sexton, M., Ceraul, S. M., Gillespie, J. J., & Azada, A. F. (2012). TolC-dependent secretion of an ankyrin repeat-containing protein of Rickettsia typhi. Journal of Bacteriology, 194(18), 4920-4932. https://doi.org/10.1128/JB.00793-12

TolC-dependent secretion of an ankyrin repeat-containing protein of Rickettsia typhi. / Kaur, Simran J.; Sayeedur Rahman, M.; Ammerman, Nicole; Beier-Sexton, Magda; Ceraul, Shane M.; Gillespie, Joseph J.; Azada, Abdu F.

In: Journal of Bacteriology, Vol. 194, No. 18, 01.09.2012, p. 4920-4932.

Research output: Contribution to journalArticle

Kaur, SJ, Sayeedur Rahman, M, Ammerman, N, Beier-Sexton, M, Ceraul, SM, Gillespie, JJ & Azada, AF 2012, 'TolC-dependent secretion of an ankyrin repeat-containing protein of Rickettsia typhi', Journal of Bacteriology, vol. 194, no. 18, pp. 4920-4932. https://doi.org/10.1128/JB.00793-12
Kaur SJ, Sayeedur Rahman M, Ammerman N, Beier-Sexton M, Ceraul SM, Gillespie JJ et al. TolC-dependent secretion of an ankyrin repeat-containing protein of Rickettsia typhi. Journal of Bacteriology. 2012 Sep 1;194(18):4920-4932. https://doi.org/10.1128/JB.00793-12
Kaur, Simran J. ; Sayeedur Rahman, M. ; Ammerman, Nicole ; Beier-Sexton, Magda ; Ceraul, Shane M. ; Gillespie, Joseph J. ; Azada, Abdu F. / TolC-dependent secretion of an ankyrin repeat-containing protein of Rickettsia typhi. In: Journal of Bacteriology. 2012 ; Vol. 194, No. 18. pp. 4920-4932.
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