Expression of the Rickettsia prowazekii pld or tlyC gene in Salmonella enterica serovar typhimurium mediates phagosomal escape

Ted Whitworth, Vsevolod Popov, Xue Jie Yu, David Walker, Donald Bouyer

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82 Citations (Scopus)

Abstract

Members of the genus Rickettsia possess the ability to invade host cells and promptly escape from phagosomal vacuoles into the host cell cytosol, thereby avoiding destruction within the endosomal pathway. The mechanism underlying rickettsial phagosomal escape remains unknown, although the genomic sequences of several rickettsial species have allowed for the identification of four genes with potential membranolytic activities (tlyA, tlyC, pat1, and pld). This study was undertaken to determine which of the selected genes of Rickettsia prowazekii mediate the escape process. Quantitative ultrastructural analyses indicated that the period of active phagosomal escape was between 30 and 50 min postinfection. Reverse transcriptase PCR analyses determined that tlyC and pld were transcribed during the period of active phagosomal escape but that tlyA and pat1 were not. The functionality of both tlyC and pld was determined by complementation studies of Salmonella, which replicates within endosomes. Complementation of Salmonella organisms with either tlyC or pld resulted in the escape of transformants from endosomal vacuoles into the host cell cytosol demonstrated by quantitative ultrastructural analyses. These data suggest a role for tlyC and pld in the process of phagosomal escape by A. prowazekii.

Original languageEnglish
Pages (from-to)6668-6673
Number of pages6
JournalInfection and Immunity
Volume73
Issue number10
DOIs
StatePublished - Oct 2005

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Rickettsia prowazekii
Salmonella enterica
Vacuoles
Salmonella
Cytosol
Genes
Rickettsia
Endosomes
Reverse Transcriptase Polymerase Chain Reaction
Demography
Serogroup

ASJC Scopus subject areas

  • Immunology

Cite this

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title = "Expression of the Rickettsia prowazekii pld or tlyC gene in Salmonella enterica serovar typhimurium mediates phagosomal escape",
abstract = "Members of the genus Rickettsia possess the ability to invade host cells and promptly escape from phagosomal vacuoles into the host cell cytosol, thereby avoiding destruction within the endosomal pathway. The mechanism underlying rickettsial phagosomal escape remains unknown, although the genomic sequences of several rickettsial species have allowed for the identification of four genes with potential membranolytic activities (tlyA, tlyC, pat1, and pld). This study was undertaken to determine which of the selected genes of Rickettsia prowazekii mediate the escape process. Quantitative ultrastructural analyses indicated that the period of active phagosomal escape was between 30 and 50 min postinfection. Reverse transcriptase PCR analyses determined that tlyC and pld were transcribed during the period of active phagosomal escape but that tlyA and pat1 were not. The functionality of both tlyC and pld was determined by complementation studies of Salmonella, which replicates within endosomes. Complementation of Salmonella organisms with either tlyC or pld resulted in the escape of transformants from endosomal vacuoles into the host cell cytosol demonstrated by quantitative ultrastructural analyses. These data suggest a role for tlyC and pld in the process of phagosomal escape by A. prowazekii.",
author = "Ted Whitworth and Vsevolod Popov and Yu, {Xue Jie} and David Walker and Donald Bouyer",
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AU - Popov, Vsevolod

AU - Yu, Xue Jie

AU - Walker, David

AU - Bouyer, Donald

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