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Human Metapneumovirus Is Capable of Entering Cells by Fusion with Endosomal Membranes

Cox, RG and Mainou, BA and Johnson, M and Hastings, AK and Schuster, JE and Dermody, TS and Williams, JV (2015) Human Metapneumovirus Is Capable of Entering Cells by Fusion with Endosomal Membranes. PLoS Pathogens, 11 (12). ISSN 1553-7366

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Abstract

© 2015 Cox et al. Human metapneumovirus (HMPV), a member of the Paramyxoviridae family, is a leading cause of lower respiratory illness. Although receptor binding is thought to initiate fusion at the plasma membrane for paramyxoviruses, the entry mechanism for HMPV is largely uncharacterized. Here we sought to determine whether HMPV initiates fusion at the plasma membrane or following internalization. To study the HMPV entry process in human bronchial epithelial (BEAS-2B) cells, we used fluorescence microscopy, an R18-dequenching fusion assay, and developed a quantitative, fluorescence microscopy assay to follow virus binding, internalization, membrane fusion, and visualize the cellular site of HMPV fusion. We found that HMPV particles are internalized into human bronchial epithelial cells before fusing with endosomes. Using chemical inhibitors and RNA interference, we determined that HMPV particles are internalized via clathrin-mediated endocytosis in a dynamin-dependent manner. HMPV fusion and productive infection are promoted by RGD-binding integrin engagement, internalization, actin polymerization, and dynamin. Further, HMPV fusion is pH-independent, although infection with rare strains is modestly inhibited by RNA interference or chemical inhibition of endosomal acidification. Thus, HMPV can enter via endocytosis, but the viral fusion machinery is not triggered by low pH. Together, our results indicate that HMPV is capable of entering host cells by multiple pathways, including membrane fusion from endosomal compartments.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Cox, RG
Mainou, BA
Johnson, Mmoj11@pitt.eduMOJ11
Hastings, AK
Schuster, JE
Dermody, TS
Williams, JV
Contributors:
ContributionContributors NameEmailPitt UsernameORCID
EditorLee, BenhurUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Date: 1 January 2015
Date Type: Publication
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Journal or Publication Title: PLoS Pathogens
Volume: 11
Number: 12
DOI or Unique Handle: 10.1371/journal.ppat.1005303
Institution: University of Pittsburgh
Schools and Programs: School of Medicine > Pediatrics
Refereed: Yes
ISSN: 1553-7366
Date Deposited: 23 Aug 2016 13:39
Last Modified: 13 Oct 2017 22:59
URI: http://d-scholarship.pitt.edu/id/eprint/28569

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