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  1. Article ; Online: Characterization and functional interrogation of the SARS-CoV-2 RNA interactome.

    Labeau, Athéna / Fery-Simonian, Luc / Lefevre-Utile, Alain / Pourcelot, Marie / Bonnet-Madin, Lucie / Soumelis, Vassili / Lotteau, Vincent / Vidalain, Pierre-Olivier / Amara, Ali / Meertens, Laurent

    Cell reports

    2022  Volume 39, Issue 4, Page(s) 110744

    Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the COVID-19 pandemic, which has led to a devastating global health crisis. The emergence of variants that escape neutralizing responses emphasizes the urgent need to ... ...

    Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the COVID-19 pandemic, which has led to a devastating global health crisis. The emergence of variants that escape neutralizing responses emphasizes the urgent need to deepen our understanding of SARS-CoV-2 biology. Using a comprehensive identification of RNA-binding proteins (RBPs) by mass spectrometry (ChIRP-MS) approach, we identify 107 high-confidence cellular factors that interact with the SARS-CoV-2 genome during infection. By systematically knocking down their expression in human lung epithelial cells, we find that the majority of the identified RBPs are SARS-CoV-2 proviral factors. In particular, we show that HNRNPA2B1, ILF3, QKI, and SFPQ interact with the SARS-CoV-2 genome and promote viral RNA amplification. Our study provides valuable resources for future investigations into the mechanisms of SARS-CoV-2 replication and the identification of host-centered antiviral therapies.
    MeSH term(s) COVID-19/genetics ; Humans ; Pandemics ; RNA, Viral/genetics ; SARS-CoV-2/genetics ; Virus Replication/genetics
    Chemical Substances RNA, Viral
    Language English
    Publishing date 2022-04-24
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2649101-1
    ISSN 2211-1247 ; 2211-1247
    ISSN (online) 2211-1247
    ISSN 2211-1247
    DOI 10.1016/j.celrep.2022.110744
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: A Genome-Wide CRISPR-Cas9 Screen Identifies the Dolichol-Phosphate Mannose Synthase Complex as a Host Dependency Factor for Dengue Virus Infection.

    Labeau, Athena / Simon-Loriere, Etienne / Hafirassou, Mohamed-Lamine / Bonnet-Madin, Lucie / Tessier, Sarah / Zamborlini, Alessia / Dupré, Thierry / Seta, Nathalie / Schwartz, Olivier / Chaix, Marie-Laure / Delaugerre, Constance / Amara, Ali / Meertens, Laurent

    Journal of virology

    2020  Volume 94, Issue 7

    Abstract: Dengue virus (DENV) is a mosquito-borne flavivirus responsible for dengue disease, a major human health concern for which no specific therapies are available. Like other viruses, DENV relies heavily on the host cellular machinery for productive infection. ...

    Abstract Dengue virus (DENV) is a mosquito-borne flavivirus responsible for dengue disease, a major human health concern for which no specific therapies are available. Like other viruses, DENV relies heavily on the host cellular machinery for productive infection. In this study, we performed a genome-wide CRISPR-Cas9 screen using haploid HAP1 cells to identify host genes important for DENV infection. We identified DPM1 and -3, two subunits of the endoplasmic reticulum (ER) resident dolichol-phosphate mannose synthase (DPMS) complex, as host dependency factors for DENV and other related flaviviruses, such as Zika virus (ZIKV). The DPMS complex catalyzes the synthesis of dolichol-phosphate mannose (DPM), which serves as mannosyl donor in pathways leading to N-glycosylation, glycosylphosphatidylinositol (GPI) anchor biosynthesis, and C- or O-mannosylation of proteins in the ER lumen. Mutation in the DXD motif of DPM1, which is essential for its catalytic activity, abolished DPMS-mediated DENV infection. Similarly, genetic ablation of ALG3, a mannosyltransferase that transfers mannose to lipid-linked oligosaccharide (LLO), rendered cells poorly susceptible to DENV. We also established that in cells deficient for DPMS activity, viral RNA amplification is hampered and truncated oligosaccharides are transferred to the viral prM and E glycoproteins, affecting their proper folding. Overall, our study provides new insights into the host-dependent mechanisms of DENV infection and supports current therapeutic approaches using glycosylation inhibitors to treat DENV infection.
    MeSH term(s) Animals ; CRISPR-Cas Systems ; Chlorocebus aethiops ; Dengue/virology ; Dengue Virus/physiology ; Endoplasmic Reticulum/metabolism ; Fibroblasts/metabolism ; Glycoproteins/metabolism ; Glycosylation ; Glycosylphosphatidylinositols/metabolism ; HEK293 Cells ; Humans ; Mannose/chemistry ; Mannosyltransferases/metabolism ; Oligosaccharides/chemistry ; RNA, Guide, CRISPR-Cas Systems/metabolism ; RNA, Viral/chemistry ; Vero Cells ; Virus Replication
    Chemical Substances Glycoproteins ; Glycosylphosphatidylinositols ; Oligosaccharides ; RNA, Guide, CRISPR-Cas Systems ; RNA, Viral ; Mannosyltransferases (EC 2.4.1.-) ; dolichyl-phosphate beta-D-mannosyltransferase (EC 2.4.1.83) ; Mannose (PHA4727WTP)
    Language English
    Publishing date 2020-03-17
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/JVI.01751-19
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: A Global Interactome Map of the Dengue Virus NS1 Identifies Virus Restriction and Dependency Host Factors.

    Hafirassou, Mohamed Lamine / Meertens, Laurent / Umaña-Diaz, Claudia / Labeau, Athena / Dejarnac, Ophelie / Bonnet-Madin, Lucie / Kümmerer, Beate M / Delaugerre, Constance / Roingeard, Philippe / Vidalain, Pierre-Olivier / Amara, Ali

    Cell reports

    2018  Volume 22, Issue 5, Page(s) 1364

    Language English
    Publishing date 2018-03-09
    Publishing country United States
    Document type Journal Article ; Published Erratum
    ZDB-ID 2649101-1
    ISSN 2211-1247 ; 2211-1247
    ISSN (online) 2211-1247
    ISSN 2211-1247
    DOI 10.1016/j.celrep.2018.01.038
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Characterization and functional interrogation of SARS-CoV-2 RNA interactome

    Labeau, Athena / Lefevre-Utile, Alain / Bonnet-Madin, Lucie / Fery-Simonian, Luc / Soumelis, Vassili / Lotteau, Vincent / Vidalain, Pierre-Olivier / Amara, Ali / Meertens, Laurent

    bioRxiv

    Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of COVID-19 pandemic, which has caused a devastating global health crisis. The emergence of highly transmissible novel viral strains that escape neutralizing responses ... ...

    Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of COVID-19 pandemic, which has caused a devastating global health crisis. The emergence of highly transmissible novel viral strains that escape neutralizing responses emphasizes the urgent need to deepen our understanding of SARS-CoV-2 biology and to develop additional therapeutic strategies. Using a comprehensive identification of RNA binding proteins (RBP) by mass spectrometry (ChIRP-M/S) approach, we identified 142 high-confidence cellular factors that bind the SARS-CoV-2 viral genome during infection. By systematically knocking down their expression in a human lung epithelial cell line, we found that the majority of the RBPs identified in our study are proviral factors that regulate SARS-CoV-2 genome replication. We showed that some of these proteins represented drug targets of interest for inhibiting SARS-CoV-2 infection. In conclusion, this study provides a comprehensive view of the SARS-CoV-2 RNA interactome during infection and highlights candidates for host-centered antiviral therapies.
    Keywords covid19
    Language English
    Publishing date 2021-03-23
    Publisher Cold Spring Harbor Laboratory
    Document type Article ; Online
    DOI 10.1101/2021.03.23.436611
    Database COVID19

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  5. Article ; Online: A Global Interactome Map of the Dengue Virus NS1 Identifies Virus Restriction and Dependency Host Factors

    Mohamed Lamine Hafirassou / Laurent Meertens / Claudia Umaña-Diaz / Athena Labeau / Ophelie Dejarnac / Lucie Bonnet-Madin / Beate M. Kümmerer / Constance Delaugerre / Philippe Roingeard / Pierre-Olivier Vidalain / Ali Amara

    Cell Reports, Vol 22, Iss 5, p

    2018  Volume 1364

    Keywords Biology (General) ; QH301-705.5
    Language English
    Publishing date 2018-01-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: FHL1 is a key player of chikungunya virus tropism and pathogenesis.

    Meertens, Laurent / Hafirassou, Mohamed Lamine / Couderc, Thérèse / Bonnet-Madin, Lucie / Kril, Vasiliya / Kümmerer, Beate M / Labeau, Athena / Brugier, Alexis / Simon-Loriere, Etienne / Burlaud-Gaillard, Julien / Doyen, Cécile / Pezzi, Laura / Goupil, Thibaud / Rafasse, Sophia / Vidalain, Pierre-Olivier / Legout, Anne Bertrand / Gueneau, Lucie / Juntas-Morales, Raul / Yaou, Rabah Ben /
    Bonne, Gisèle / de Lamballerie, Xavier / Benkirane, Monsef / Roingeard, Philippe / Delaugerre, Constance / Lecuit, Marc / Amara, Ali

    Comptes rendus biologies

    2021  Volume 343, Issue 4, Page(s) 79–89

    Abstract: Chikungunya is an infectious disease caused by the chikungunya virus (CHIKV), an alphavirus transmitted to humans by Aedes mosquitoes, and for which there is no licensed vaccine nor antiviral treatments. By using a loss-of-function genetic screen, we ... ...

    Abstract Chikungunya is an infectious disease caused by the chikungunya virus (CHIKV), an alphavirus transmitted to humans by Aedes mosquitoes, and for which there is no licensed vaccine nor antiviral treatments. By using a loss-of-function genetic screen, we have recently identified the FHL1 protein as an essential host factor for CHIKV tropism and pathogenesis. FHL1 is highly expressed in muscles cells and fibroblasts, the main CHIKV-target cells. FHL1 interacts with the viral protein nsP3 and plays a critical role in CHIKV genome amplification. Experiments in vivo performed in FHL1-deficient mice have shown that these animals are resistant to infection and do not develop muscular lesions. Altogether these observations, published in the journal Nature [1], show that FHL1 is a key host factor for CHIKV pathogenesis and identify the interaction between FHL1 and nsP3 as a promising target for the development of new antiviral strategies.
    MeSH term(s) Animals ; Chikungunya Fever ; Chikungunya virus/genetics ; Intracellular Signaling Peptides and Proteins ; LIM Domain Proteins ; Mice ; Muscle Proteins ; Tropism ; Viral Nonstructural Proteins ; Virus Replication
    Chemical Substances Fhl1 protein, mouse ; Intracellular Signaling Peptides and Proteins ; LIM Domain Proteins ; Muscle Proteins ; Viral Nonstructural Proteins
    Language English
    Publishing date 2021-04-21
    Publishing country France
    Document type Journal Article
    ZDB-ID 2072863-3
    ISSN 1768-3238 ; 1631-0691
    ISSN (online) 1768-3238
    ISSN 1631-0691
    DOI 10.5802/crbiol.40
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: A Global Interactome Map of the Dengue Virus NS1 Identifies Virus Restriction and Dependency Host Factors.

    Hafirassou, Mohamed Lamine / Meertens, Laurent / Umaña-Diaz, Claudia / Labeau, Athena / Dejarnac, Ophelie / Bonnet-Madin, Lucie / Kümmerer, Beate M / Delaugerre, Constance / Roingeard, Philippe / Vidalain, Pierre-Olivier / Amara, Ali

    Cell reports

    2017  Volume 21, Issue 13, Page(s) 3900–3913

    Abstract: Dengue virus (DENV) infections cause the most prevalent mosquito-borne viral disease worldwide, for which no therapies are available. DENV encodes seven non-structural (NS) proteins that co-assemble and recruit poorly characterized host factors to form ... ...

    Abstract Dengue virus (DENV) infections cause the most prevalent mosquito-borne viral disease worldwide, for which no therapies are available. DENV encodes seven non-structural (NS) proteins that co-assemble and recruit poorly characterized host factors to form the DENV replication complex essential for viral infection. Here, we provide a global proteomic analysis of the human host factors that interact with the DENV NS1 protein. Combined with a functional RNAi screen, this study reveals a comprehensive network of host cellular processes involved in DENV infection and identifies DENV host restriction and dependency factors. We highlight an important role of RACK1 and the chaperonin TRiC (CCT) and oligosaccharyltransferase (OST) complexes during DENV replication. We further show that the OST complex mediates NS1 and NS4B glycosylation, and pharmacological inhibition of its N-glycosylation function strongly impairs DENV infection. In conclusion, our study provides a global interactome of the DENV NS1 and identifies host factors targetable for antiviral therapies.
    MeSH term(s) Dengue/virology ; Dengue Virus/metabolism ; Glycosylation ; HEK293 Cells ; HeLa Cells ; Host-Pathogen Interactions ; Humans ; Multiprotein Complexes/metabolism ; Neoplasm Proteins/metabolism ; Protein Interaction Maps ; RNA, Small Interfering/metabolism ; Receptors for Activated C Kinase/metabolism ; Viral Nonstructural Proteins/metabolism ; Virus Replication
    Chemical Substances Multiprotein Complexes ; Neoplasm Proteins ; RACK1 protein, human ; RNA, Small Interfering ; Receptors for Activated C Kinase ; Viral Nonstructural Proteins
    Language English
    Publishing date 2017-12-27
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2649101-1
    ISSN 2211-1247 ; 2211-1247
    ISSN (online) 2211-1247
    ISSN 2211-1247
    DOI 10.1016/j.celrep.2017.11.094
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: A Global Interactome Map of the Dengue Virus NS1 Identifies Virus Restriction and Dependency Host Factors

    Mohamed Lamine Hafirassou / Laurent Meertens / Claudia Umaña-Diaz / Athena Labeau / Ophelie Dejarnac / Lucie Bonnet-Madin / Beate M. Kümmerer / Constance Delaugerre / Philippe Roingeard / Pierre-Olivier Vidalain / Ali Amara

    Cell Reports, Vol 21, Iss 13, Pp 3900-

    2017  Volume 3913

    Abstract: Dengue virus (DENV) infections cause the most prevalent mosquito-borne viral disease worldwide, for which no therapies are available. DENV encodes seven non-structural (NS) proteins that co-assemble and recruit poorly characterized host factors to form ... ...

    Abstract Dengue virus (DENV) infections cause the most prevalent mosquito-borne viral disease worldwide, for which no therapies are available. DENV encodes seven non-structural (NS) proteins that co-assemble and recruit poorly characterized host factors to form the DENV replication complex essential for viral infection. Here, we provide a global proteomic analysis of the human host factors that interact with the DENV NS1 protein. Combined with a functional RNAi screen, this study reveals a comprehensive network of host cellular processes involved in DENV infection and identifies DENV host restriction and dependency factors. We highlight an important role of RACK1 and the chaperonin TRiC (CCT) and oligosaccharyltransferase (OST) complexes during DENV replication. We further show that the OST complex mediates NS1 and NS4B glycosylation, and pharmacological inhibition of its N-glycosylation function strongly impairs DENV infection. In conclusion, our study provides a global interactome of the DENV NS1 and identifies host factors targetable for antiviral therapies.
    Keywords Biology (General) ; QH301-705.5
    Subject code 570
    Language English
    Publishing date 2017-12-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  9. Article ; Online: Axl Mediates ZIKA Virus Entry in Human Glial Cells and Modulates Innate Immune Responses

    Laurent Meertens / Athena Labeau / Ophelie Dejarnac / Sara Cipriani / Laura Sinigaglia / Lucie Bonnet-Madin / Tifenn Le Charpentier / Mohamed Lamine Hafirassou / Alessia Zamborlini / Van-Mai Cao-Lormeau / Muriel Coulpier / Dorothée Missé / Nolwenn Jouvenet / Ray Tabibiazar / Pierre Gressens / Olivier Schwartz / Ali Amara

    Cell Reports, Vol 18, Iss 2, Pp 324-

    2017  Volume 333

    Abstract: Summary: ZIKA virus (ZIKV) is an emerging pathogen responsible for neurological disorders and congenital microcephaly. However, the molecular basis for ZIKV neurotropism remains poorly understood. Here, we show that Axl is expressed in human microglia ... ...

    Abstract Summary: ZIKA virus (ZIKV) is an emerging pathogen responsible for neurological disorders and congenital microcephaly. However, the molecular basis for ZIKV neurotropism remains poorly understood. Here, we show that Axl is expressed in human microglia and astrocytes in the developing brain and that it mediates ZIKV infection of glial cells. Axl-mediated ZIKV entry requires the Axl ligand Gas6, which bridges ZIKV particles to glial cells. Following binding, ZIKV is internalized through clathrin-mediated endocytosis and traffics to Rab5+ endosomes to establish productive infection. During entry, the ZIKV/Gas6 complex activates Axl kinase activity, which downmodulates interferon signaling and facilitates infection. ZIKV infection of human glial cells is inhibited by MYD1, an engineered Axl decoy receptor, and by the Axl kinase inhibitor R428. Our results highlight the dual role of Axl during ZIKV infection of glial cells: promoting viral entry and modulating innate immune responses. Therefore, inhibiting Axl function may represent a potential target for future antiviral therapies. : ZIKA virus (ZIKV) is responsible for congenital microcephaly. Meertens et al. show that the Axl receptor is expressed in microglia and astrocytes in the human developing brain. They also highlight the dual role of Axl during infection, which promotes viral entry and dampens innate immune responses in glial cells.
    Keywords Biology (General) ; QH301-705.5
    Subject code 570
    Language English
    Publishing date 2017-01-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  10. Article ; Online: FHL1 is a major host factor for chikungunya virus infection.

    Meertens, Laurent / Hafirassou, Mohamed Lamine / Couderc, Thérèse / Bonnet-Madin, Lucie / Kril, Vasiliya / Kümmerer, Beate M / Labeau, Athena / Brugier, Alexis / Simon-Loriere, Etienne / Burlaud-Gaillard, Julien / Doyen, Cécile / Pezzi, Laura / Goupil, Thibaud / Rafasse, Sophia / Vidalain, Pierre-Olivier / Bertrand-Legout, Anne / Gueneau, Lucie / Juntas-Morales, Raul / Ben Yaou, Rabah /
    Bonne, Gisèle / de Lamballerie, Xavier / Benkirane, Monsef / Roingeard, Philippe / Delaugerre, Constance / Lecuit, Marc / Amara, Ali

    Nature

    2019  Volume 574, Issue 7777, Page(s) 259–263

    Abstract: Chikungunya virus (CHIKV) is a re-emerging alphavirus that is transmitted to humans by mosquito bites and causes musculoskeletal and joint ... ...

    Abstract Chikungunya virus (CHIKV) is a re-emerging alphavirus that is transmitted to humans by mosquito bites and causes musculoskeletal and joint pain
    MeSH term(s) Animals ; Cells, Cultured ; Chikungunya Fever/drug therapy ; Chikungunya Fever/virology ; Chikungunya virus/drug effects ; Chikungunya virus/genetics ; Chikungunya virus/growth & development ; Chikungunya virus/pathogenicity ; Female ; Fibroblasts/virology ; HEK293 Cells ; Host-Derived Cellular Factors/genetics ; Host-Derived Cellular Factors/metabolism ; Host-Pathogen Interactions ; Humans ; Intracellular Signaling Peptides and Proteins/deficiency ; Intracellular Signaling Peptides and Proteins/genetics ; Intracellular Signaling Peptides and Proteins/metabolism ; LIM Domain Proteins/deficiency ; LIM Domain Proteins/genetics ; LIM Domain Proteins/metabolism ; Male ; Mice ; Muscle Proteins/deficiency ; Muscle Proteins/genetics ; Muscle Proteins/metabolism ; Myoblasts/virology ; O'nyong-nyong Virus/growth & development ; O'nyong-nyong Virus/pathogenicity ; Protein Binding ; RNA, Viral/biosynthesis ; Viral Nonstructural Proteins/genetics ; Viral Nonstructural Proteins/metabolism ; Virus Replication
    Chemical Substances FHL1 protein, human ; Host-Derived Cellular Factors ; Intracellular Signaling Peptides and Proteins ; LIM Domain Proteins ; Muscle Proteins ; RNA, Viral ; Viral Nonstructural Proteins ; nsp3 protein, alphavirus
    Language English
    Publishing date 2019-09-25
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 120714-3
    ISSN 1476-4687 ; 0028-0836
    ISSN (online) 1476-4687
    ISSN 0028-0836
    DOI 10.1038/s41586-019-1578-4
    Database MEDical Literature Analysis and Retrieval System OnLINE

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