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Identification of candidate host cell factors required for actin-based motility of Burkholderia pseudomallei

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Original languageEnglish
Pages (from-to)4675-4685
JournalJournal Of Proteome Research
Volume15
Issue number12
Early online date14 Nov 2016
DOIs
Publication statusE-pub ahead of print - 14 Nov 2016

Abstract

Intracellular actin-based motility of the melioidosis pathogen Burkholderia pseudomallei requires the bacterial factor BimA. Located at one pole of the bacterium, BimA recruits and polymerises cellular actin to promote bacterial motility within and between cells. Here we describe an affinity approach coupled with Mass Spectrometry to identify cellular proteins recruited to BimA-expressing bacteria under conditions that promote actin polymerisation. We identified a group of cellular proteins that are recruited to the B. pseudomallei surface in a BimA-dependent manner, a subset of which were independently validated with specific antisera including the ubiquitous scaffold protein Ras GTPase-activating-like protein (IQGAP1). IQGAP1 integrates several key cellular signalling pathways including those involved in actin dynamics and has been shown to be involved in the adhesion of attaching and effacing Escherichia coli to infected cells and invasion of host cells by Salmonella enterica serovar Typhimurium. Whilst a direct interaction between BimA and IQGAP1 could not be detected using either conventional pulldown or yeast two hybrid techniques, confocal microscopy revealed that IQGAP1 is recruited to B. pseudomallei actin tails in infected cells and siRNA-mediated knockdown highlighted a role for this protein in controlling the length and actin density of B. pseudomallei actin tails.

    Research areas

  • Burkholderia pseudomallei, melioidosis, intracellular bacterial pathogen, actin-based motility, BimA, IQGAP1

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