Dataset Information

GPI-Anchored Ligand-BioID2-Tagging System Identifies proteins Mediating Zika Virus Entry

ABSTRACT: Identification of host factors facilitating pathogen entry is critical for preventing infectious diseases. Here, we report a tagging system consisting of a viral receptor-binding protein (RBP) linked to BioID2, which is expressed on the cell surface via a GPI anchor. Using VSV or Zika virus (ZIKV) RBP, the system (BioID2- RBP(V)-GPI; BioID2-RBP(Z)-GPI) faithfully identifies LDLR and AXL, the receptors of VSV and ZIKV, respectively. Being GPI-anchored is essential for the probe to function properly. Furthermore, BioID2-RBP(Z)-GPI expressed in human neuronal progenitor cells identifies galectin-1 on cell surface pivotal for ZIKV entry. This conclusion is further supported by antibody blocking and galectin-1 silencing in A549 and mouse neural cells. Importantly, Lgas1-/- mice are significantly more resistant to ZIKV infection than Lgas1+/+ littermates are, having significantly lower virus titers and fewer pathologies in various organs. This tagging system may have broad applications for identifying protein-protein interactions on the cell surface

ORGANISM(S): Homo Sapiens

SUBMITTER: Chaoyang Li

PROVIDER: PXD037307 | iProX | Wed Oct 12 00:00:00 BST 2022

REPOSITORIES: iProX

ACCESS DATA

Publications

GPI-anchored ligand-BioID2-tagging system identifies Galectin-1 mediating Zika virus entry.

Gao Shan-Shan SS Shi Run R Sun Jing J Tang Yanhong Y Zheng Zhenhua Z Li Jing-Feng JF Li Huan H Zhang Jie J Leng Qibin Q Xu Jiang J Chen Xinwen X Zhao Jincun J Sy Man-Sun MS Feng Liqiang L Li Chaoyang C

iScience 20221102 12

Identification of host factors facilitating pathogen entry is critical for preventing infectious diseases. Here, we report a tagging system consisting of a viral receptor-binding protein (RBP) linked to BioID2, which is expressed on the cell surface via a GPI anchor. Using VSV or Zika virus (ZIKV) RBP, the system (BioID2- RBP(V)-GPI; BioID2-RBP(Z)-GPI) faithfully identifies LDLR and AXL, the receptors of VSV and ZIKV, respectively. Being GPI-anchored is essential for the probe to function proper ...[more]

PMID: 36404916

Similar Datasets

Project description:Previous microarray data indicated a significant increase in interferon (IFN) stimulated gene expression in cells exposed to conditioned medium from primary human trophoblasts compared to non conditioned medium. Here we want to compare the effects of conditioned media, non conditioned media, and non conditioned media spiked with 100 units of IFN beta in wild type HT1080 cells as well as in HT1080 cells deficient in STAT1 (a key signaling molecular in the IFN pathway). Interferon stimulated genes (ISGs) target viruses at various stages of their infectious life cycles, including at the earliest stage of viral entry. Here we identify ArfGAP with dual pleckstrin homology (PH) domains 2 (ADAP2) as a gene upregulated by type I IFN treatment in a STAT1-dependent manner. ADAP2 functions as a GTPase-activating protein (GAP) for Arf6 and binds to phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3) and PI(3,4)P2. We show that overexpression of ADAP2 suppresses dengue virus (DENV) and vesicular stomatitis virus (VSV) infection in an Arf6 GAP activity-dependent manner, while exerting no effect on coxsackievirus B (CVB) or Sendai virus (SeV) replication. We further show that ADAP2 expression induces macropinocytosis and that ADAP2 strongly associates with actin-enriched membrane ruffles and with Rab8a- and LAMP1-, but not EEA1- or Rab7-, positive vesicles. Utilizing two techniques—light-sensitive neutral red (NR)-containing DENV and fluorescence assays for virus internalization—we show that ADAP2 primarily restricts DENV infection at the stage of virion entry and/or intracellular trafficking and that incoming DENV and VSV particles associate with ADAP2 during their entry. Taken together, this study identifies ADAP2 as an ISG that exerts antiviral effects against RNA viruses by altering Arf6-mediated trafficking to disrupt viral entry. compare the cells exposed to CM and the cells exposed to NC media spiked with IFN to the control NC exposed cells, in wild type (2f-TGH) vs Stat -/- cells (HT1080U3A)

Dataset Information

GPI-Anchored Ligand-BioID2-Tagging System Identifies proteins Mediating Zika Virus Entry

Publications

GPI-anchored ligand-BioID2-tagging system identifies Galectin-1 mediating Zika virus entry.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure