Extracellular secretion of the virulence plasmid-encoded ADP-ribosyltransferase SpvB in Salmonella

Hideo Gotoh, Nobuhiko Okada, Yungi Kim, Kouya Shiraishi, Naoko Hirami, Takeshi Haneda, Ai Kurita, Yuji Kikuchi, Hirofumi Danbara

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Nontyphoid Salmonella enterica requires the plasmid-encoded spv genes to establish successful systemic infection in experimental animals. The SpvB virulence-associated protein has recently been shown to contain the ADP-ribosyltransferase domain. SpvB ADP-ribosilates actin and depolymerizes actin filaments when expressed in cultured epithelial cells. However, spontaneous secretion or release of SpvB has not been observed under in vitro growth conditions. In the present study we investigated the secretion of SpvB from Salmonella using in vitro and in vivo assay systems. We showed that SpvB is secreted into supernatant from Salmonella strains that contain the cloned spvB gene on a plasmid when they grew in intracellular salts medium (ISM), a minimal medium mimicing the intracellular iron concentrations of eukaryotic cells. A series of mutant SpvB proteins revealed that an N-terminal region of SpvB located at amino acids 1-229 was sufficient to promote secretion into extracellular milieu. Confocal immunofluorescence microscopy also demonstrated efficient localization of the N-terminal domain of SpvB1-360 tagged with biotinylated peptide within infected host cell cytosol but not truncated SpvB1-179 fusion protein. In addition, mutations that inactivate genes within Salmonella pathogenicity island 1 or Salmonella pathogenicity island 2 that encode type III secretion systems (TTSS) could secrete the SpvB protein into the culture medium. These results indicate that SpvB protein is transported from the bacteria and into the host cytoplasm independent of TTSS.

Original languageEnglish
Pages (from-to)227-238
Number of pages12
JournalMicrobial Pathogenesis
Volume34
Issue number5
DOIs
Publication statusPublished - 2003 May 1
Externally publishedYes

Fingerprint

ADP Ribose Transferases
Salmonella
Virulence
Plasmids
Genomic Islands
Proteins
Genes
Salmonella enterica
Eukaryotic Cells
Mutant Proteins
Actin Cytoskeleton
Fluorescence Microscopy
Confocal Microscopy
Cytosol
Culture Media
Cultured Cells
Cytoplasm
Iron
Salts
Epithelial Cells

Keywords

  • ADP-ribosyltransferase
  • Salmonella
  • Secretion
  • SpvB

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Extracellular secretion of the virulence plasmid-encoded ADP-ribosyltransferase SpvB in Salmonella. / Gotoh, Hideo; Okada, Nobuhiko; Kim, Yungi; Shiraishi, Kouya; Hirami, Naoko; Haneda, Takeshi; Kurita, Ai; Kikuchi, Yuji; Danbara, Hirofumi.

In: Microbial Pathogenesis, Vol. 34, No. 5, 01.05.2003, p. 227-238.

Research output: Contribution to journalArticle

Gotoh, H, Okada, N, Kim, Y, Shiraishi, K, Hirami, N, Haneda, T, Kurita, A, Kikuchi, Y & Danbara, H 2003, 'Extracellular secretion of the virulence plasmid-encoded ADP-ribosyltransferase SpvB in Salmonella', Microbial Pathogenesis, vol. 34, no. 5, pp. 227-238. https://doi.org/10.1016/S0882-4010(03)00034-2
Gotoh, Hideo ; Okada, Nobuhiko ; Kim, Yungi ; Shiraishi, Kouya ; Hirami, Naoko ; Haneda, Takeshi ; Kurita, Ai ; Kikuchi, Yuji ; Danbara, Hirofumi. / Extracellular secretion of the virulence plasmid-encoded ADP-ribosyltransferase SpvB in Salmonella. In: Microbial Pathogenesis. 2003 ; Vol. 34, No. 5. pp. 227-238.
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