Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
A Role for the VPS Retromer in Brucella Intracellular Replication Revealed by Genomewide siRNA Screening
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..
Show others and affiliations
2019 (English)In: mSphere, ISSN 2379-5042, Vol. 4, no 3, article id e00380-19Article in journal (Refereed) Published
Abstract [en]

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 degradative lysosomal pathway and to establish the ER-derived replicative BCV (rBCV) have already been identified, the global understanding of this highly coordinated process is still partial, and many factors remain unknown. To gain deeper insight into these fundamental questions, we performed a genomewide RNA interference (RNAi) screen aiming at discovering novel host factors involved in the Brucella intracellular cycle. We identified 425 host proteins that contribute to Brucella cellular entry, intracellular trafficking, and replication. Together, this study sheds light on previously unknown host pathways required for the Brucella infection cycle and highlights the VPS retromer components as critical factors for the establishment of the Brucella intracellular replicative niche.

Place, publisher, year, edition, pages
American Society for Microbiology , 2019. Vol. 4, no 3, article id e00380-19
Keywords [en]
Brucella, RAB7A, VPS retromer, VPS26, VPS35, replicative niche, siRNA
National Category
Cell Biology
Identifiers
URN: urn:nbn:se:kth:diva-255585DOI: 10.1128/mSphere.00380-19ISI: 000475754600003PubMedID: 31243080Scopus ID: 2-s2.0-85068819548OAI: oai:DiVA.org:kth-255585DiVA, id: diva2:1341081
Note

QC 20190807

Available from: 2019-08-07 Created: 2019-08-07 Last updated: 2019-09-16Bibliographically approved

Open Access in DiVA

fulltext(6386 kB)43 downloads
File information
File name FULLTEXT01.pdfFile size 6386 kBChecksum SHA-512
af6c4e866d30b8d012f980419dd41cf8264571d9d8f697b5bff7ed56124cd5d3e9c2e1cf272efee84feed898a60ca96647ef6e39033a137cd74d5868243624d7
Type fulltextMimetype application/pdf

Other links

Publisher's full textPubMedScopus

Search in DiVA

By author/editor
Smith, Kevin
By organisation
Science for Life Laboratory, SciLifeLabSchool of Computer Science and Communication (CSC)
Cell Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 43 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 57 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf