A Role for the VPS Retromer in Brucella Intracellular Replication Revealed by Genomewide siRNA Screening
KTH, Science for Life Laboratory, SciLifeLab ; KTH, Skolan för datavetenskap och kommunikation (CSC) ; Univ Basel, Biozentrum, Basel, Switzerland.;Novartis Pharma Schweiz AG, Rotkreuz, Switzerland. ; Univ Basel, Biozentrum, Basel, Switzerland.;Nanyang Technol Univ, Lee Kong China Sch Med, Singapore, Singapore. ; Univ Basel, Biozentrum, Basel, Switzerland. ; Univ Basel, Biozentrum, Basel, Switzerland ; Univ Basel, Biozentrum, Basel, Switzerland.;BioDataAnalysis GmbH, Munich, Germany. ; Univ Basel, Biozentrum, Basel, Switzerland.;Univ Hosp Basel, Basel, Switzerland. ; American Society for Microbiology, 2019
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Zugriff:
Brucella, the agent causing brucellosis, is a major zoonotic pathogen with worldwide distribution. Brucella resides and replicates inside infected host cells in membrane-bound compartments called Brucella-containing vacuoles (BCVs). Following uptake, Brucella resides in endosomal BCVs (eBCVs) that gradually mature from early to late endosomal features. Through a poorly understood process that is key to the intracellular lifestyle of Brucella, the eBCV escapes fusion with lysosomes by transitioning to the replicative BCV (rBCV), a replicative niche directly connected to the endoplasmic reticulum (ER). Despite the notion that this complex intracellular lifestyle must depend on a multitude of host factors, a holistic view on which of these components control Brucella cell entry, trafficking, and replication is still missing. Here we used a systematic cell-based small interfering RNA (siRNA) knockdown screen in HeLa cells infected with Brucella abortus and identified 425 components of the human infectome for Brucella infection. These include multiple components of pathways involved in central processes such as the cell cycle, actin cytoskeleton dynamics, or vesicular trafficking. Using assays for pathogen entry, knockdown complementation, and colocalization at single-cell resolution, we identified the requirement of the VPS retromer for Brucella to escape the lysosomal degradative pathway and to establish its intracellular replicative niche. We thus validated the VPS retromer as a novel host factor critical for Brucella intracellular trafficking. Further, our genomewide data shed light on the interplay between central host processes and the biogenesis of the Brucella replicative niche. IMPORTANCE With >300,000 new cases of human brucellosis annually, Brucella is regarded as one of the most important zoonotic bacterial pathogens worldwide. The agent causing brucellosis resides inside host cells within vacuoles termed Brucella-containing vacuoles (BCVs). Although a few host components required to escape the ...
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A Role for the VPS Retromer in Brucella Intracellular Replication Revealed by Genomewide siRNA Screening
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Autor/in / Beteiligte Person: | Casanova, Alain ; Low, Shyan Huey ; Quebatte, Maxime ; Sedzicki, Jaroslaw ; Tschon, Therese ; Ketterer, Maren ; Smith, Kevin ; Emmenlauer, Mario ; Ben-Tekaya, Houchaima ; Dehio, Christoph |
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Veröffentlichung: | KTH, Science for Life Laboratory, SciLifeLab ; KTH, Skolan för datavetenskap och kommunikation (CSC) ; Univ Basel, Biozentrum, Basel, Switzerland.;Novartis Pharma Schweiz AG, Rotkreuz, Switzerland. ; Univ Basel, Biozentrum, Basel, Switzerland.;Nanyang Technol Univ, Lee Kong China Sch Med, Singapore, Singapore. ; Univ Basel, Biozentrum, Basel, Switzerland. ; Univ Basel, Biozentrum, Basel, Switzerland ; Univ Basel, Biozentrum, Basel, Switzerland.;BioDataAnalysis GmbH, Munich, Germany. ; Univ Basel, Biozentrum, Basel, Switzerland.;Univ Hosp Basel, Basel, Switzerland. ; American Society for Microbiology, 2019 |
Medientyp: | academicJournal |
DOI: | 10.1128/mSphere.00380-19 |
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