title: Burkholderia pseudomalleitype III secreted protein BipC: role in actin modulation and translocation activities required for the bacterial intracellular lifecycle.
abstract: Melioidosis, an infection caused by the facultative intracellular pathogenBurkholderia pseudomallei, has been classified as an emerging disease with the number of patients steadily increasing at an alarming rate.B. pseudomalleipossess various virulence determinants that allow them to invade the host and evade the host immune response, such as the type III secretion systems (TTSS). The products of this specialized secretion system are particularly important for theB. pseudomalleiinfection. Lacking in one or more components of the TTSS demonstrated different degrees of defects in the intracellular lifecycle ofB. pseudomallei. Further understanding the functional roles of proteins involved inB. pseudomalleiTTSS will enable us to dissect the enigma ofB. pseudomallei-host cell interaction. In this study, BipC (a translocator), which was previously reported to be involved in the pathogenesis ofB. pseudomallei, was further characterized using the bioinformatics and molecular approaches. ThebipCgene, coding for a putative invasive protein, was first PCR amplified fromB. pseudomalleiK96243 genomic DNA and cloned into an expression vector for overexpression inEscherichia coli. The soluble protein was subsequently purified and assayed for actin polymerization and depolymerization. BipC was verified to subvert the host actin dynamics as demonstrated by the capability to polymerize actinin vitro. Homology modeling was also attempted to predict the structure of BipC. Overall, our findings identified that the protein encoded by thebipCgene plays a role as an effector involved in the actin binding activity to facilitate internalization ofB. pseudomalleiinto the host cells.
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya , Kuala Lumpur , Malaysia.
authors.forename: Wen Tyng
authors.initials: WT
authors.lastname: Kang
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya , Kuala Lumpur , Malaysia.
authors.forename: Kumutha Malar
authors.initials: KM
authors.lastname: Vellasamy
authors.affiliation: Antimicrobials, Singapore Eye Research Institute (SERI) , Singapore , Singapore.
authors.forename: Lakshminarayanan
authors.initials: L
authors.lastname: Rajamani
authors.affiliation: Antimicrobials, Singapore Eye Research Institute (SERI) , Singapore , Singapore.
authors.forename: Roger W
authors.initials: RW
authors.lastname: Beuerman
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya , Kuala Lumpur , Malaysia.
authors.forename: Jamuna
authors.initials: J
authors.lastname: Vadivelu
keywords: Actin dynamics; Burkholderia pseudomallei; Effector protein; Intracellular lifecycle; Protein expression and purification; Type III secretion system