The Brucella effector BspL targets the ER-associated degradation (ERAD) pathway and delays bacterial egress from infected cells

Jean-Baptiste Luizet; Julie Raymond; Thais Lourdes Santos Lacerda; Emeline BARBIEUX; Stanimir Kambarev; Magali Bonici; Frédérique Lembo; KEVIN WILLEMART; Jean-Paul Borg; Jean Celli; Francine C. A. Gérard; Eric MURAILLE; Jean-Pierre Gorvel; Suzana P Salcedo

doi:10.1073/pnas.2105324118

The Brucella effector BspL targets the ER-associated degradation (ERAD) pathway and delays bacterial egress from infected cells

Jean-Baptiste Luizet
, Julie Raymond
, Thais Lourdes Santos Lacerda
, Emeline BARBIEUX
, Stanimir Kambarev
, Magali Bonici
, Frédérique Lembo
, KEVIN WILLEMART
, Jean-Paul Borg
, Jean Celli
, Francine C. A. Gérard
, Eric MURAILLE
, Jean-Pierre Gorvel
, Suzana P Salcedo

Research output: Contribution to journal › Article › peer-review

Abstract

Perturbation of the endoplasmic reticulum (ER), a central organelle of the cell, can have critical consequences for cellular homeostasis. An elaborate surveillance system known as ER quality control ensures that cells can respond and adapt to stress via the unfolded protein response (UPR) and that only correctly assembled proteins reach their destination. Interestingly, several bacterial pathogens hijack the ER to establish an infection. However, it remains poorly understood how bacterial pathogens exploit ER quality-control functions to complete their intracellular cycle. Brucella spp. replicate extensively within an ER-derived niche, which evolves into specialized vacuoles suited for exit from infected cells. Here we present Brucella-secreted protein L (BspL), a Brucella abortus effector that interacts with Herp, a central component of the ERassociated degradation (ERAD) machinery. We found that BspL enhances ERAD at the late stages of the infection. BspL targeting of Herp and ERAD allows tight control of the kinetics of autophagic Brucella-containing vacuole formation, delaying the last step of its intracellular cycle and cell-to-cell spread. This study highlights a mechanism by which a bacterial pathogen hijacks ERAD components for fine regulation of its intracellular trafficking.

Original language	English
Article number	e2105324118
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	118
Issue number	32
DOIs	https://doi.org/10.1073/pnas.2105324118
Publication status	Published - 10 Aug 2021

Funding

We thank Linda Hendershot (St. Jude's Children's Research Hospital) for sending us the pcDNA-TCR\u03B1 and for all the help with setting up the endoplasmic reticulum-associated degradation assay and discussion of the results; Thomas Rutkowski (University of Iowa) for the cDNA encoding the Null Hong Kong mutant of \u03B11-antitrypsin; Ren\u00E9e Tsolis (University of California, Davis) for help constructing TEM1-VceC and HA-VceC; Thomas Henry (Centre International de Recherche en Infectologie, Lyon, France) for the immortalized cell line of bone marrow-derived macrophages; Steve Garvis, Amandine Blanco, and Arthur Louche for the critical reading of the manuscript; and Peter Cresswell and Susan Mitchell (Yale University School of Medicine) for sending us the endoplasmic reticulum-associated degradation reporter cell line and all the advice despite all the challenges imposed by the pandemic. All microscopy imaging was carried out at the PLATIM microscopy facility. The two-hybrid screening was hosted by the Marseille Proteomics platform (J.-P.B. and F.L.) supported by Institut Paoli- Calmettes, Infrastructures Biologie Sant\u00E9 et Agronomie, Aix-Marseille University, Cancerop\u00F4le PACA, and the R\u00E9gion Sud Provence-Alpes-C\u00F4te d'Azur. J.-P.B. is a scholar of Institut Universitaire de France. This work was funded by the ERA-Net Pathogenomics CELLPATH Grant Agriculture and Natural Resources (ANR) 2010-PATH-006, the FINOVI foundation under a Young Researcher Starting grant, ANR NUCPATH Grant ANR-15-CE15-0011, and the ANR charm-Ed Grant ANR-18-CE15-0003 (all obtained by S.P.S.). J.-B.L. was supported by a doctoral contract from the R\u00E9gion Rh\u00F4nes-Alpes ARC1 Sant\u00E9. An INSERM staff scientist contract supports S.P.S. The original submission of this manuscript was published on BiorXiv and is part of the PhD thesis manuscript of J.-B.L., \"Host Cell modulation by Brucella effectors,\" Universit\u00E9 de Lyon, France, 2019. ACKNOWLEDGMENTS. We thank Linda Hendershot (St. Jude\u2019s Children\u2019s Research Hospital) for sending us the pcDNA-TCR\u03B1 and for all the help with setting up the endoplasmic reticulum-associated degradation assay and discussion of the results; Thomas Rutkowski (University of Iowa) for the cDNA encoding the Null Hong Kong mutant of \u03B11-antitrypsin; Ren\u00E9e Tsolis (University of California, Davis) for help constructing TEM1-VceC and HA-VceC; Thomas Henry (Centre International de Recherche en Infectologie, Lyon, France) for the immortalized cell line of bone marrow-derived macrophages; Steve Garvis, Amandine Blanco, and Arthur Louche for the critical reading of the manuscript; and Peter Cresswell and Susan Mitchell (Yale University School of Medicine) for sending us the endoplasmic reticulum-associated degradation reporter cell line and all the advice despite all the challenges imposed by the pandemic. All microscopy imaging was carried out at the PLATIM microscopy facility. The two-hybrid screening was hosted by the Marseille Proteomics platform (J.-P.B. and F.L.) supported by Institut Paoli-Calmettes, Infrastructures Biologie Sant\u00E9 et Agronomie, Aix-Marseille University, Cancerop\u00F4le PACA, and the R\u00E9gion Sud Provence-Alpes-C\u00F4te d\u2019Azur. J.-P.B. is a scholar of Institut Universitaire de France. This work was funded by the ERA-Net Pathogenomics CELLPATH Grant Agriculture and Natural Resources (ANR) 2010-PATH-006, the FINOVI foundation under a Young Researcher Starting grant, ANR NUCPATH Grant ANR-15-CE15-0011, and the ANR charm-Ed Grant ANR-18-CE15-0003 (all obtained by S.P.S.). J.-B.L. was supported by a doctoral contract from the R\u00E9gion Rh\u00F4nes-Alpes ARC1 Sant\u00E9. An INSERM staff scientist contract supports S.P.S. The original submission of this manuscript was published on BiorXiv and is part of the PhD thesis manuscript of J.-B.L., \u201CHost Cell modulation by Brucella effectors,\u201D Universit\u00E9 de Lyon, France, 2019.

Funders	Funder number
Institut National de la Santé et de la Recherche Médicale
Institut Paoli-Calmettes
Aix-Marseille Université
Institut Paoli- Calmettes
Université de Lyon
Institut Universitaire de France
Infrastructures en Biologie Santé et Agronomie
University of Iowa
Yale University
Thomas Rutkowski
Région Rhônes-Alpes ARC1 Santé
Conseil Régional Provence-Alpes-Côte d'Azur	2010-PATH-006
Fondation Innovations en Infectiologie	ANR-18-CE15-0003
Agence Nationale de la Recherche	ANR-15-CE15-0011

Keywords

Brucella
ERAD
Herp
Trafficking

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10.1073/pnas.2105324118

Cite this

Luizet, J.-B., Raymond, J., Lourdes Santos Lacerda, T., BARBIEUX, E., Kambarev, S., Bonici, M., Lembo, F., WILLEMART, KEVIN., Borg, J.-P., Celli, J., Gérard, F. C. A., MURAILLE, E., Gorvel, J.-P., & Salcedo, S. P. (2021). The Brucella effector BspL targets the ER-associated degradation (ERAD) pathway and delays bacterial egress from infected cells. Proceedings of the National Academy of Sciences of the United States of America, 118(32), Article e2105324118. https://doi.org/10.1073/pnas.2105324118

@article{2e0f3fddef2a4c198a3ff9aab4507ede,

title = "The Brucella effector BspL targets the ER-associated degradation (ERAD) pathway and delays bacterial egress from infected cells",

abstract = "Perturbation of the endoplasmic reticulum (ER), a central organelle of the cell, can have critical consequences for cellular homeostasis. An elaborate surveillance system known as ER quality control ensures that cells can respond and adapt to stress via the unfolded protein response (UPR) and that only correctly assembled proteins reach their destination. Interestingly, several bacterial pathogens hijack the ER to establish an infection. However, it remains poorly understood how bacterial pathogens exploit ER quality-control functions to complete their intracellular cycle. Brucella spp. replicate extensively within an ER-derived niche, which evolves into specialized vacuoles suited for exit from infected cells. Here we present Brucella-secreted protein L (BspL), a Brucella abortus effector that interacts with Herp, a central component of the ERassociated degradation (ERAD) machinery. We found that BspL enhances ERAD at the late stages of the infection. BspL targeting of Herp and ERAD allows tight control of the kinetics of autophagic Brucella-containing vacuole formation, delaying the last step of its intracellular cycle and cell-to-cell spread. This study highlights a mechanism by which a bacterial pathogen hijacks ERAD components for fine regulation of its intracellular trafficking.",

keywords = "Brucella, ERAD, Herp, Trafficking",

author = "Jean-Baptiste Luizet and Julie Raymond and \{Lourdes Santos Lacerda\}, Thais and Emeline BARBIEUX and Stanimir Kambarev and Magali Bonici and Fr{\'e}d{\'e}rique Lembo and KEVIN WILLEMART and Jean-Paul Borg and Jean Celli and G{\'e}rard, \{Francine C. A.\} and Eric MURAILLE and Jean-Pierre Gorvel and Salcedo, \{Suzana P\}",

note = "Funding Information: We thank Linda Hendershot (St. Jude's Children's Research Hospital) for sending us the pcDNA-TCRα and for all the help with setting up the endoplasmic reticulum-associated degradation assay and discussion of the results; Thomas Rutkowski (University of Iowa) for the cDNA encoding the Null Hong Kong mutant of α1-antitrypsin; Ren{\'e}e Tsolis (University of California, Davis) for help constructing TEM1-VceC and HA-VceC; Thomas Henry (Centre International de Recherche en Infectologie, Lyon, France) for the immortalized cell line of bone marrow-derived macrophages; Steve Garvis, Amandine Blanco, and Arthur Louche for the critical reading of the manuscript; and Peter Cresswell and Susan Mitchell (Yale University School of Medicine) for sending us the endoplasmic reticulum-associated degradation reporter cell line and all the advice despite all the challenges imposed by the pandemic. All microscopy imaging was carried out at the PLATIM microscopy facility. The two-hybrid screening was hosted by the Marseille Proteomics platform (J.-P.B. and F.L.) supported by Institut Paoli- Calmettes, Infrastructures Biologie Sant{\'e} et Agronomie, Aix-Marseille University, Cancerop{\^o}le PACA, and the R{\'e}gion Sud Provence-Alpes-C{\^o}te d'Azur. J.-P.B. is a scholar of Institut Universitaire de France. This work was funded by the ERA-Net Pathogenomics CELLPATH Grant Agriculture and Natural Resources (ANR) 2010-PATH-006, the FINOVI foundation under a Young Researcher Starting grant, ANR NUCPATH Grant ANR-15-CE15-0011, and the ANR charm-Ed Grant ANR-18-CE15-0003 (all obtained by S.P.S.). J.-B.L. was supported by a doctoral contract from the R{\'e}gion Rh{\^o}nes-Alpes ARC1 Sant{\'e}. An INSERM staff scientist contract supports S.P.S. The original submission of this manuscript was published on BiorXiv and is part of the PhD thesis manuscript of J.-B.L., {"}Host Cell modulation by Brucella effectors,{"} Universit{\'e} de Lyon, France, 2019. Funding Information: ACKNOWLEDGMENTS. We thank Linda Hendershot (St. Jude{\textquoteright}s Children{\textquoteright}s Research Hospital) for sending us the pcDNA-TCRα and for all the help with setting up the endoplasmic reticulum-associated degradation assay and discussion of the results; Thomas Rutkowski (University of Iowa) for the cDNA encoding the Null Hong Kong mutant of α1-antitrypsin; Ren{\'e}e Tsolis (University of California, Davis) for help constructing TEM1-VceC and HA-VceC; Thomas Henry (Centre International de Recherche en Infectologie, Lyon, France) for the immortalized cell line of bone marrow-derived macrophages; Steve Garvis, Amandine Blanco, and Arthur Louche for the critical reading of the manuscript; and Peter Cresswell and Susan Mitchell (Yale University School of Medicine) for sending us the endoplasmic reticulum-associated degradation reporter cell line and all the advice despite all the challenges imposed by the pandemic. All microscopy imaging was carried out at the PLATIM microscopy facility. The two-hybrid screening was hosted by the Marseille Proteomics platform (J.-P.B. and F.L.) supported by Institut Paoli-Calmettes, Infrastructures Biologie Sant{\'e} et Agronomie, Aix-Marseille University, Cancerop{\^o}le PACA, and the R{\'e}gion Sud Provence-Alpes-C{\^o}te d{\textquoteright}Azur. J.-P.B. is a scholar of Institut Universitaire de France. This work was funded by the ERA-Net Pathogenomics CELLPATH Grant Agriculture and Natural Resources (ANR) 2010-PATH-006, the FINOVI foundation under a Young Researcher Starting grant, ANR NUCPATH Grant ANR-15-CE15-0011, and the ANR charm-Ed Grant ANR-18-CE15-0003 (all obtained by S.P.S.). J.-B.L. was supported by a doctoral contract from the R{\'e}gion Rh{\^o}nes-Alpes ARC1 Sant{\'e}. An INSERM staff scientist contract supports S.P.S. The original submission of this manuscript was published on BiorXiv and is part of the PhD thesis manuscript of J.-B.L., “Host Cell modulation by Brucella effectors,” Universit{\'e} de Lyon, France, 2019. Publisher Copyright: {\textcopyright} 2021 National Academy of Sciences. All rights reserved.",

year = "2021",

month = aug,

day = "10",

doi = "10.1073/pnas.2105324118",

language = "English",

volume = "118",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "National Academy of Sciences",

number = "32",

}

Luizet, J-B, Raymond, J, Lourdes Santos Lacerda, T, BARBIEUX, E, Kambarev, S, Bonici, M, Lembo, F, WILLEMART, KEVIN, Borg, J-P, Celli, J, Gérard, FCA, MURAILLE, E, Gorvel, J-P & Salcedo, SP 2021, 'The Brucella effector BspL targets the ER-associated degradation (ERAD) pathway and delays bacterial egress from infected cells', Proceedings of the National Academy of Sciences of the United States of America, vol. 118, no. 32, e2105324118. https://doi.org/10.1073/pnas.2105324118

The Brucella effector BspL targets the ER-associated degradation (ERAD) pathway and delays bacterial egress from infected cells. / Luizet, Jean-Baptiste; Raymond, Julie; Lourdes Santos Lacerda, Thais et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 118, No. 32, e2105324118, 10.08.2021.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - The Brucella effector BspL targets the ER-associated degradation (ERAD) pathway and delays bacterial egress from infected cells

AU - Luizet, Jean-Baptiste

AU - Raymond, Julie

AU - Lourdes Santos Lacerda, Thais

AU - BARBIEUX, Emeline

AU - Kambarev, Stanimir

AU - Bonici, Magali

AU - Lembo, Frédérique

AU - WILLEMART, KEVIN

AU - Borg, Jean-Paul

AU - Celli, Jean

AU - Gérard, Francine C. A.

AU - MURAILLE, Eric

AU - Gorvel, Jean-Pierre

AU - Salcedo, Suzana P

N1 - Funding Information: We thank Linda Hendershot (St. Jude's Children's Research Hospital) for sending us the pcDNA-TCRα and for all the help with setting up the endoplasmic reticulum-associated degradation assay and discussion of the results; Thomas Rutkowski (University of Iowa) for the cDNA encoding the Null Hong Kong mutant of α1-antitrypsin; Renée Tsolis (University of California, Davis) for help constructing TEM1-VceC and HA-VceC; Thomas Henry (Centre International de Recherche en Infectologie, Lyon, France) for the immortalized cell line of bone marrow-derived macrophages; Steve Garvis, Amandine Blanco, and Arthur Louche for the critical reading of the manuscript; and Peter Cresswell and Susan Mitchell (Yale University School of Medicine) for sending us the endoplasmic reticulum-associated degradation reporter cell line and all the advice despite all the challenges imposed by the pandemic. All microscopy imaging was carried out at the PLATIM microscopy facility. The two-hybrid screening was hosted by the Marseille Proteomics platform (J.-P.B. and F.L.) supported by Institut Paoli- Calmettes, Infrastructures Biologie Santé et Agronomie, Aix-Marseille University, Canceropôle PACA, and the Région Sud Provence-Alpes-Côte d'Azur. J.-P.B. is a scholar of Institut Universitaire de France. This work was funded by the ERA-Net Pathogenomics CELLPATH Grant Agriculture and Natural Resources (ANR) 2010-PATH-006, the FINOVI foundation under a Young Researcher Starting grant, ANR NUCPATH Grant ANR-15-CE15-0011, and the ANR charm-Ed Grant ANR-18-CE15-0003 (all obtained by S.P.S.). J.-B.L. was supported by a doctoral contract from the Région Rhônes-Alpes ARC1 Santé. An INSERM staff scientist contract supports S.P.S. The original submission of this manuscript was published on BiorXiv and is part of the PhD thesis manuscript of J.-B.L., "Host Cell modulation by Brucella effectors," Université de Lyon, France, 2019. Funding Information: ACKNOWLEDGMENTS. We thank Linda Hendershot (St. Jude’s Children’s Research Hospital) for sending us the pcDNA-TCRα and for all the help with setting up the endoplasmic reticulum-associated degradation assay and discussion of the results; Thomas Rutkowski (University of Iowa) for the cDNA encoding the Null Hong Kong mutant of α1-antitrypsin; Renée Tsolis (University of California, Davis) for help constructing TEM1-VceC and HA-VceC; Thomas Henry (Centre International de Recherche en Infectologie, Lyon, France) for the immortalized cell line of bone marrow-derived macrophages; Steve Garvis, Amandine Blanco, and Arthur Louche for the critical reading of the manuscript; and Peter Cresswell and Susan Mitchell (Yale University School of Medicine) for sending us the endoplasmic reticulum-associated degradation reporter cell line and all the advice despite all the challenges imposed by the pandemic. All microscopy imaging was carried out at the PLATIM microscopy facility. The two-hybrid screening was hosted by the Marseille Proteomics platform (J.-P.B. and F.L.) supported by Institut Paoli-Calmettes, Infrastructures Biologie Santé et Agronomie, Aix-Marseille University, Canceropôle PACA, and the Région Sud Provence-Alpes-Côte d’Azur. J.-P.B. is a scholar of Institut Universitaire de France. This work was funded by the ERA-Net Pathogenomics CELLPATH Grant Agriculture and Natural Resources (ANR) 2010-PATH-006, the FINOVI foundation under a Young Researcher Starting grant, ANR NUCPATH Grant ANR-15-CE15-0011, and the ANR charm-Ed Grant ANR-18-CE15-0003 (all obtained by S.P.S.). J.-B.L. was supported by a doctoral contract from the Région Rhônes-Alpes ARC1 Santé. An INSERM staff scientist contract supports S.P.S. The original submission of this manuscript was published on BiorXiv and is part of the PhD thesis manuscript of J.-B.L., “Host Cell modulation by Brucella effectors,” Université de Lyon, France, 2019. Publisher Copyright: © 2021 National Academy of Sciences. All rights reserved.

PY - 2021/8/10

Y1 - 2021/8/10

N2 - Perturbation of the endoplasmic reticulum (ER), a central organelle of the cell, can have critical consequences for cellular homeostasis. An elaborate surveillance system known as ER quality control ensures that cells can respond and adapt to stress via the unfolded protein response (UPR) and that only correctly assembled proteins reach their destination. Interestingly, several bacterial pathogens hijack the ER to establish an infection. However, it remains poorly understood how bacterial pathogens exploit ER quality-control functions to complete their intracellular cycle. Brucella spp. replicate extensively within an ER-derived niche, which evolves into specialized vacuoles suited for exit from infected cells. Here we present Brucella-secreted protein L (BspL), a Brucella abortus effector that interacts with Herp, a central component of the ERassociated degradation (ERAD) machinery. We found that BspL enhances ERAD at the late stages of the infection. BspL targeting of Herp and ERAD allows tight control of the kinetics of autophagic Brucella-containing vacuole formation, delaying the last step of its intracellular cycle and cell-to-cell spread. This study highlights a mechanism by which a bacterial pathogen hijacks ERAD components for fine regulation of its intracellular trafficking.

AB - Perturbation of the endoplasmic reticulum (ER), a central organelle of the cell, can have critical consequences for cellular homeostasis. An elaborate surveillance system known as ER quality control ensures that cells can respond and adapt to stress via the unfolded protein response (UPR) and that only correctly assembled proteins reach their destination. Interestingly, several bacterial pathogens hijack the ER to establish an infection. However, it remains poorly understood how bacterial pathogens exploit ER quality-control functions to complete their intracellular cycle. Brucella spp. replicate extensively within an ER-derived niche, which evolves into specialized vacuoles suited for exit from infected cells. Here we present Brucella-secreted protein L (BspL), a Brucella abortus effector that interacts with Herp, a central component of the ERassociated degradation (ERAD) machinery. We found that BspL enhances ERAD at the late stages of the infection. BspL targeting of Herp and ERAD allows tight control of the kinetics of autophagic Brucella-containing vacuole formation, delaying the last step of its intracellular cycle and cell-to-cell spread. This study highlights a mechanism by which a bacterial pathogen hijacks ERAD components for fine regulation of its intracellular trafficking.

KW - Brucella

KW - ERAD

KW - Herp

KW - Trafficking

UR - https://www.scopus.com/pages/publications/85112652810

U2 - 10.1073/pnas.2105324118

DO - 10.1073/pnas.2105324118

M3 - Article

SN - 0027-8424

VL - 118

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 32

M1 - e2105324118

ER -

The Brucella effector BspL targets the ER-associated degradation (ERAD) pathway and delays bacterial egress from infected cells

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