Molecular characterization of a functional type VI secretion system from a clinical isolate of Aeromonas hydrophila

Giovanni Suarez, Johanna C. Sierra, Jian Sha, Shaofei Wang, Tatiana E. Erova, Amin A. Fadl, Sheri M. Foltz, Amy J. Horneman, Ashok Chopra

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Abstract

Our laboratory recently molecularly characterized the type II secretion system (T2SS)-associated cytotoxic enterotoxin (Act) and the T3SS-secreted AexU effector from a diarrheal isolate SSU of Aeromonas hydrophila. The role of these toxin proteins in the pathogenesis of A. hydrophila infections was subsequently delineated in in vitro and in vivo models. In this study, we characterized the new type VI secretion system (T6SS) from isolate SSU of A. hydrophila and demonstrated its role in bacterial virulence. Study of the role of T6SS in bacterial virulence is in its infancy, and there are, accordingly, only limited, recent reports directed toward a better understanding its role in bacterial pathogenesis. We have provided evidence that the virulence-associated secretion (vas) genes vasH (Sigma 54-dependent transcriptional regulator) and vasK (encoding protein of unknown function) are essential for expression of the genes encoding the T6SS and/or they constituted important components of the T6SS. Deletion of the vasH gene prevented expression of the potential translocon hemolysin coregulated protein (Hcp) encoding gene from bacteria, while the vasK gene deletion prevented secretion but not translocation of Hcp into host cells. The secretion of Hcp was independent of the T3SS and the flagellar system. We demonstrated that secreted Hcp could bind to the murine RAW 264.7 macrophages from outside, in addition to its ability to be translocated into host cells. Further, the vasH and vasK mutants were less toxic to murine macrophages and human epithelial HeLa cells, and these mutants were more efficiently phagocytosed by macrophages. We also provided evidence that the expression of the hcp gene in the HeLa cell resulted in apoptosis of the host cells. Finally, the vasH and vasK mutants of A. hydrophila were less virulent in a septicemic mouse model of infection, and animals immunized with recombinant Hcp were protected from subsequent challenge with the wild-type (WT) bacterium. In addition, mice infected with the WT A. hydrophila had circulating antibodies to Hcp, indicating an important role of T6SS in the pathogenesis of A. hydrophila infections. Taken together, we have characterized the T6SS from Aeromonas for the first time and provided new features of this secretion system not yet known for other pathogens.

Original languageEnglish
Pages (from-to)344-361
Number of pages18
JournalMicrobial Pathogenesis
Volume44
Issue number4
DOIs
StatePublished - Apr 2008

Fingerprint

Aeromonas hydrophila
Hemolysin Proteins
Virulence
Macrophages
HeLa Cells
Infection
Bacteria
Gene Expression
Aeromonas
Poisons
Enterotoxins
Essential Genes
Gene Deletion
Phagocytosis
Recombinant Proteins
Genes
Type VI Secretion Systems
Proteins
Animal Models
Epithelial Cells

Keywords

  • Aeromonas hydrophila
  • Isogenic mutants
  • Murine model of toxicity
  • Translocation and secretion of effectors
  • Type VI secretion system

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Molecular characterization of a functional type VI secretion system from a clinical isolate of Aeromonas hydrophila. / Suarez, Giovanni; Sierra, Johanna C.; Sha, Jian; Wang, Shaofei; Erova, Tatiana E.; Fadl, Amin A.; Foltz, Sheri M.; Horneman, Amy J.; Chopra, Ashok.

In: Microbial Pathogenesis, Vol. 44, No. 4, 04.2008, p. 344-361.

Research output: Contribution to journalArticle

Suarez, Giovanni ; Sierra, Johanna C. ; Sha, Jian ; Wang, Shaofei ; Erova, Tatiana E. ; Fadl, Amin A. ; Foltz, Sheri M. ; Horneman, Amy J. ; Chopra, Ashok. / Molecular characterization of a functional type VI secretion system from a clinical isolate of Aeromonas hydrophila. In: Microbial Pathogenesis. 2008 ; Vol. 44, No. 4. pp. 344-361.
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