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dc.contributor.author
Krzyzaniak, Magdalena A.
dc.contributor.author
Zumstein, Michael T.
dc.contributor.author
Gerez, Juan A.
dc.contributor.author
Picotti, Paola
dc.contributor.author
Helenius, Ari
dc.date.accessioned
2018-11-27T11:16:49Z
dc.date.available
2017-06-10T16:20:44Z
dc.date.available
2018-11-06T17:28:38Z
dc.date.available
2018-11-27T11:16:49Z
dc.date.issued
2013-04-11
dc.identifier.other
10.1371/journal.ppat.1003309
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/66141
dc.identifier.doi
10.3929/ethz-b-000066141
dc.description.abstract
Respiratory Syncytial Virus (RSV) is a highly pathogenic member of the Paramyxoviridae that causes severe respiratory tract infections. Reports in the literature have indicated that to infect cells the incoming viruses either fuse their envelope directly with the plasma membrane or exploit clathrin-mediated endocytosis. To study the entry process in human tissue culture cells (HeLa, A549), we used fluorescence microscopy and developed quantitative, FACS-based assays to follow virus binding to cells, endocytosis, intracellular trafficking, membrane fusion, and infection. A variety of perturbants were employed to characterize the cellular processes involved. We found that immediately after binding to cells RSV activated a signaling cascade involving the EGF receptor, Cdc42, PAK1, and downstream effectors. This led to a series of dramatic actin rearrangements; the cells rounded up, plasma membrane blebs were formed, and there was a significant increase in fluid uptake. If these effects were inhibited using compounds targeting Na+/H+ exchangers, myosin II, PAK1, and other factors, no infection was observed. The RSV was rapidly and efficiently internalized by an actin-dependent process that had all hallmarks of macropinocytosis. Rather than fusing with the plasma membrane, the viruses thus entered Rab5-positive, fluid-filled macropinosomes, and fused with the membranes of these on the average 50 min after internalization. Rab5 was required for infection. To find an explanation for the endocytosis requirement, which is unusual among paramyxoviruses, we analyzed the fusion protein, F, and could show that, although already cleaved by a furin family protease once, it underwent a second, critical proteolytic cleavage after internalization. This cleavage by a furin-like protease removed a small peptide from the F1 subunits, and made the virus infectious.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
Public Library Science (PLOS)
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/3.0/
dc.title
Host Cell Entry of Respiratory Syncytial Virus Involves Macropinocytosis Followed by Proteolytic Activation of the F Protein
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 3.0 Unported
ethz.journal.title
PLoS pathogens
ethz.journal.volume
9
en_US
ethz.journal.issue
4
en_US
ethz.pages.start
e1003309
en_US
ethz.size
19 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.identifier.nebis
005409548
ethz.publication.place
Lawrence, KS
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02538 - Institut für Molekulare Systembiologie / Institute for Molecular Systems Biology::03927 - Picotti, Paola / Picotti, Paola
en_US
ethz.leitzahl
03495 - Helenius, Ari
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02538 - Institut für Molekulare Systembiologie / Institute for Molecular Systems Biology::03927 - Picotti, Paola / Picotti, Paola
ethz.leitzahl.certified
03495 - Helenius, Ari
ethz.date.deposited
2017-06-10T16:21:20Z
ethz.source
ECIT
ethz.identifier.importid
imp5936508a8de2a23700
ethz.ecitpid
pub:105478
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2017-07-26T18:44:44Z
ethz.rosetta.lastUpdated
2018-11-27T11:17:00Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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