A role for the VPS retromer in Brucella intracellular replication revealed by genomewide siRNA screening
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 g...
| Main Authors: | , , , , , , , , , |
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| Format: | Journal Article |
| Language: | English |
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2020
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| Online Access: | https://hdl.handle.net/10356/142436 |
| _version_ | 1811693886636032000 |
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| author | Casanova, Alain Low, Shyan Huey Québatte, Maxime Sedzick, Jaroslaw Tschon, Therese Ketterer, Maren Smith, Kevin Emmenlauer, Mario Ben-Tekaya, Houchaima Dehio, Christoph |
| author2 | Lee Kong Chian School of Medicine (LKCMedicine) |
| author_facet | Lee Kong Chian School of Medicine (LKCMedicine) Casanova, Alain Low, Shyan Huey Québatte, Maxime Sedzick, Jaroslaw Tschon, Therese Ketterer, Maren Smith, Kevin Emmenlauer, Mario Ben-Tekaya, Houchaima Dehio, Christoph |
| author_sort | Casanova, Alain |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T06:58:48Z |
| format | Journal Article |
| id | ntu-10356/142436 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T06:58:48Z |
| publishDate | 2020 |
| record_format | dspace |
| spelling | ntu-10356/1424362020-11-01T05:26:27Z A role for the VPS retromer in Brucella intracellular replication revealed by genomewide siRNA screening Casanova, Alain Low, Shyan Huey Québatte, Maxime Sedzick, Jaroslaw Tschon, Therese Ketterer, Maren Smith, Kevin Emmenlauer, Mario Ben-Tekaya, Houchaima Dehio, Christoph Lee Kong Chian School of Medicine (LKCMedicine) Science::Medicine Brucella RAB7A 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. Published version 2020-06-22T05:15:43Z 2020-06-22T05:15:43Z 2019 Journal Article Casanova, A., Low, S. H., Québatte, M., Sedzicki, J., Tschon, T., Ketterer, M., . . . Dehio, C. (2019). A role for the VPS retromer in Brucella intracellular replication revealed by genomewide siRNA screening. mSphere, 4(3), e00380-19-. doi:10.1128/MSPHERE.00380-19 2379-5042 https://hdl.handle.net/10356/142436 10.1128/MSPHERE.00380-19 4 2-s2.0-85068819548 3 4 en mSphere © 2019 Casanova et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. application/pdf |
| spellingShingle | Science::Medicine Brucella RAB7A Casanova, Alain Low, Shyan Huey Québatte, Maxime Sedzick, Jaroslaw Tschon, Therese Ketterer, Maren Smith, Kevin Emmenlauer, Mario Ben-Tekaya, Houchaima Dehio, Christoph A role for the VPS retromer in Brucella intracellular replication revealed by genomewide siRNA screening |
| title | A role for the VPS retromer in Brucella intracellular replication revealed by genomewide siRNA screening |
| title_full | A role for the VPS retromer in Brucella intracellular replication revealed by genomewide siRNA screening |
| title_fullStr | A role for the VPS retromer in Brucella intracellular replication revealed by genomewide siRNA screening |
| title_full_unstemmed | A role for the VPS retromer in Brucella intracellular replication revealed by genomewide siRNA screening |
| title_short | A role for the VPS retromer in Brucella intracellular replication revealed by genomewide siRNA screening |
| title_sort | role for the vps retromer in brucella intracellular replication revealed by genomewide sirna screening |
| topic | Science::Medicine Brucella RAB7A |
| url | https://hdl.handle.net/10356/142436 |
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