Proximity labelling of internalizing influenza A viruses reveals a role for neogenin in virus uptake.

Influenza A virus (IAV) is a respiratory pathogen of global concern. Entry of most IAVs is mediated by binding of viral hemagglutinin to cellular sialic acid, facilitating virus attachment. A subsequent interaction with a surface receptor(s) triggers viral uptake. Although multiple host factors invo...

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Main Authors: Milagros Sempere Borau, Victor G Gisbert, Josephine von Kempis, Laura M Arroyo-Fernández, Samira Schiefer, David Alsteens, Silke Stertz
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-07-01
Series:PLoS Pathogens
Online Access:https://doi.org/10.1371/journal.ppat.1013338
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author Milagros Sempere Borau
Victor G Gisbert
Josephine von Kempis
Laura M Arroyo-Fernández
Samira Schiefer
David Alsteens
Silke Stertz
author_facet Milagros Sempere Borau
Victor G Gisbert
Josephine von Kempis
Laura M Arroyo-Fernández
Samira Schiefer
David Alsteens
Silke Stertz
author_sort Milagros Sempere Borau
collection DOAJ
description Influenza A virus (IAV) is a respiratory pathogen of global concern. Entry of most IAVs is mediated by binding of viral hemagglutinin to cellular sialic acid, facilitating virus attachment. A subsequent interaction with a surface receptor(s) triggers viral uptake. Although multiple host factors involved in viral entry are known, the identity of these receptors remains unclear. Here, we utilized proximity labelling to acquire the interactome of epsin 1, an adaptor protein utilized by IAV for clathrin-mediated endocytosis, during virus internalization to identify them. We uncover neogenin (Neo1), a member of the immunoglobulin superfamily expressed in primary human airway cultures, as a potential epsin 1 interactor and virus receptor candidate. Knockdown of Neo1 led to a reduction in replication of H1N1, H2N2 and H5N1 IAVs in primary and immortalized lung cells. Moreover, human recombinant Neo1 was found to bind IAV with a KD of 21 ± 14 nM by atomic force microscopy and Neo1 could co-localize with incoming IAV at early times post-infection, as well as affect viral entry. As Neo1 can interact with IAV and its depletion impairs IAV entry, this study reveals its potential as an IAV internalization receptor.
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issn 1553-7366
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spelling doaj-art-6516bcb95c2b474b9cf23af8202df9402025-08-20T02:39:48ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742025-07-01217e101333810.1371/journal.ppat.1013338Proximity labelling of internalizing influenza A viruses reveals a role for neogenin in virus uptake.Milagros Sempere BorauVictor G GisbertJosephine von KempisLaura M Arroyo-FernándezSamira SchieferDavid AlsteensSilke StertzInfluenza A virus (IAV) is a respiratory pathogen of global concern. Entry of most IAVs is mediated by binding of viral hemagglutinin to cellular sialic acid, facilitating virus attachment. A subsequent interaction with a surface receptor(s) triggers viral uptake. Although multiple host factors involved in viral entry are known, the identity of these receptors remains unclear. Here, we utilized proximity labelling to acquire the interactome of epsin 1, an adaptor protein utilized by IAV for clathrin-mediated endocytosis, during virus internalization to identify them. We uncover neogenin (Neo1), a member of the immunoglobulin superfamily expressed in primary human airway cultures, as a potential epsin 1 interactor and virus receptor candidate. Knockdown of Neo1 led to a reduction in replication of H1N1, H2N2 and H5N1 IAVs in primary and immortalized lung cells. Moreover, human recombinant Neo1 was found to bind IAV with a KD of 21 ± 14 nM by atomic force microscopy and Neo1 could co-localize with incoming IAV at early times post-infection, as well as affect viral entry. As Neo1 can interact with IAV and its depletion impairs IAV entry, this study reveals its potential as an IAV internalization receptor.https://doi.org/10.1371/journal.ppat.1013338
spellingShingle Milagros Sempere Borau
Victor G Gisbert
Josephine von Kempis
Laura M Arroyo-Fernández
Samira Schiefer
David Alsteens
Silke Stertz
Proximity labelling of internalizing influenza A viruses reveals a role for neogenin in virus uptake.
PLoS Pathogens
title Proximity labelling of internalizing influenza A viruses reveals a role for neogenin in virus uptake.
title_full Proximity labelling of internalizing influenza A viruses reveals a role for neogenin in virus uptake.
title_fullStr Proximity labelling of internalizing influenza A viruses reveals a role for neogenin in virus uptake.
title_full_unstemmed Proximity labelling of internalizing influenza A viruses reveals a role for neogenin in virus uptake.
title_short Proximity labelling of internalizing influenza A viruses reveals a role for neogenin in virus uptake.
title_sort proximity labelling of internalizing influenza a viruses reveals a role for neogenin in virus uptake
url https://doi.org/10.1371/journal.ppat.1013338
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