Biogenesis of Yersinia pestis PsaA in recombinant attenuated Salmonella Typhimurium vaccine (RASV) strain

Ascención Torres-Escobar, María Dolores Juárez-Rodríguez, Roy Curtiss

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Yersinia pestis PsaA is an adhesin important for the establishment of bacterial infection. PsaA synthesis requires the products of the psaEFABC genes. Here, by prediction analysis, we identified a PsaA signal sequence with two signal peptidase (SPase) cleavage sites, type-I and type-II (SPase-I and SPase-II). By Edman degradation and site-directed mutagenesis, the precise site for one of these Spase-I PsaA cleavage sites was located between alanine and serine at positions 31 and 32, respectively. Yersinia pestis psaA expression and the role of the PsaB and PsaC proteins were evaluated in recombinant attenuated Salmonella Typhimurium vaccine strains. PsaA was detected in total extracts as a major 15-kDa (mature) and 18-kDa (unprocessed) protein bands. PsaA synthesis was not altered by a ΔA31-ΔS32 double-deletion mutation. In contrast, the synthesis of PsaA (ΔA31-ΔS32) in Y. pestis and delivery to the supernatant was decreased. Otherwise, substitution of the amino acid cysteine at position 26 by valine involved in the SPase-II cleavage site did not show any effect on the secretion of PsaA in Salmonella and Yersinia. These results help clarify the secretion pathway of PsaA for the possible development of vaccines against Y. pestis.

Original languageEnglish (US)
Pages (from-to)106-113
Number of pages8
JournalFEMS Microbiology Letters
Volume302
Issue number2
DOIs
StatePublished - Jan 2010

Fingerprint

Salmonella Vaccines
Yersinia pestis
Attenuated Vaccines
Salmonella typhimurium
Yersinia
Sequence Deletion
Secretory Pathway
Valine
Amino Acid Substitution
Protein Sorting Signals
Site-Directed Mutagenesis
Bacterial Infections
Salmonella
Alanine
Serine
Cysteine
Vaccines
type I signal peptidase
Genes
signal peptidase II

Keywords

  • Chaperone protein
  • Signal peptidase
  • Usher protein
  • Yersinia enterocolitica
  • Yersinia pseudotuberculosis

ASJC Scopus subject areas

  • Microbiology
  • Genetics
  • Molecular Biology

Cite this

Biogenesis of Yersinia pestis PsaA in recombinant attenuated Salmonella Typhimurium vaccine (RASV) strain. / Torres-Escobar, Ascención; Juárez-Rodríguez, María Dolores; Curtiss, Roy.

In: FEMS Microbiology Letters, Vol. 302, No. 2, 01.2010, p. 106-113.

Research output: Contribution to journalArticle

Torres-Escobar, Ascención ; Juárez-Rodríguez, María Dolores ; Curtiss, Roy. / Biogenesis of Yersinia pestis PsaA in recombinant attenuated Salmonella Typhimurium vaccine (RASV) strain. In: FEMS Microbiology Letters. 2010 ; Vol. 302, No. 2. pp. 106-113.
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