title: Glycoprotein B cleavage is important for murid herpesvirus 4 to infect myeloid cells.
abstract: Glycoprotein B (gB) is a conserved herpesvirus virion component implicated in membrane fusion. As with many-but not all-herpesviruses, the gB of murid herpesvirus 4 (MuHV-4) is cleaved into disulfide-linked subunits, apparently by furin. Preventing gB cleavage for some herpesviruses causes minor infection deficits in vitro, but what the cleavage contributes to host colonization has been unclear. To address this, we mutated the furin cleavage site (R-R-K-R) of the MuHV-4 gB. Abolishing gB cleavage did not affect its expression levels, glycosylation, or antigenic conformation. In vitro, mutant viruses entered fibroblasts and epithelial cells normally but had a significant entry deficit in myeloid cells such as macrophages and bone marrow-derived dendritic cells. The deficit in myeloid cells was not due to reduced virion binding or endocytosis, suggesting that gB cleavage promotes infection at a postendocytic entry step, presumably viral membrane fusion. In vivo, viruses lacking gB cleavage showed reduced lytic spread in the lungs. Alveolar epithelial cell infection was normal, but alveolar macrophage infection was significantly reduced. Normal long-term latency in lymphoid tissue was established nonetheless.
authors.affiliation: Division of Virology, Department of Pathology, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Glauser
authors.forename: Ricardo
authors.initials: R
authors.lastname: Milho
authors.forename: Bruno
authors.initials: B
authors.lastname: Frederico
authors.forename: Janet S
authors.initials: JS
authors.lastname: May
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Kratz
authors.forename: Laurent
authors.initials: L
authors.lastname: Gillet
authors.forename: Philip G
authors.initials: PG
authors.lastname: Stevenson
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000378:metabolism / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000378:metabolism / Q000821:virology; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D045683:Furin / Q000378:metabolism; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008168:Lung / Q000378:metabolism / Q000821:virology*; D016676:Macrophages, Alveolar / Q000378:metabolism / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D022423:Myeloid Cells / Q000821:virology*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018144:Rhadinovirus / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion; D014779:Virus Replication
chemical_list: D057134:Antibodies, Neutralizing; D006023:Glycoproteins; D012333:RNA, Messenger; D014759:Viral Envelope Proteins; D045683:Furin
references.citation: J Virol. 2000 Aug;74(15):6964-74
references.pmid: 10888635
references.citation: Curr Opin Struct Biol. 2009 Apr;19(2):189-96
references.pmid: 19356922
references.citation: Nat Struct Mol Biol. 2008 Oct;15(10):1024-30
references.pmid: 18776902
references.citation: J Gen Virol. 2009 Oct;90(Pt 10):2317-2330
references.pmid: 19605591
references.citation: J Virol. 2002 Feb;76(3):1252-64
references.pmid: 11773401
references.citation: J Gen Virol. 2007 Jul;88(Pt 7):1859-1865
references.pmid: 17554016
references.citation: PLoS Pathog. 2011 Nov;7(11):e1002346
references.pmid: 22102809
references.citation: J Gen Virol. 2006 Dec;87(Pt 12):3515-3527
references.pmid: 17098966
references.citation: J Gen Virol. 2012 Jun;93(Pt 6):1316-1327
references.pmid: 22377583
references.citation: Curr Opin Microbiol. 2006 Aug;9(4):423-9
references.pmid: 16814597
references.citation: J Gen Virol. 1992 Sep;73 ( Pt 9):2347-56
references.pmid: 1328491
references.citation: J Virol. 2003 Jul;77(13):7696-701
references.pmid: 12805472
references.citation: J Virol. 1999 Feb;73(2):1075-9
references.pmid: 9882308
references.citation: J Virol. 2005 Mar;79(6):3459-67
references.pmid: 15731240
references.citation: J Gen Virol. 2009 Mar;90(Pt 3):591-595
references.pmid: 19218203
references.citation: J Gen Virol. 2008 Jun;89(Pt 6):1352-1363
references.pmid: 18474550
references.citation: J Gen Virol. 1996 Apr;77 ( Pt 4):627-30
references.pmid: 8627250
references.citation: J Virol. 1999 Nov;73(11):9650-4
references.pmid: 10516078
references.citation: J Gen Virol. 1992 Dec;73 ( Pt 12):3275-9
references.pmid: 1469366
references.citation: PLoS One. 2007 Jun 27;2(6):e560
references.pmid: 17593961
references.citation: J Virol. 2009 Aug;83(15):7495-506
references.pmid: 19474103
references.citation: J Gen Virol. 2011 Sep;92(Pt 9):2020-2033
references.pmid: 21593277
references.citation: J Lipid Res. 2011 Aug;52(8):1542-50
references.pmid: 21572087
references.citation: PLoS Pathog. 2012 Sep;8(9):e1002935
references.pmid: 23028329
references.citation: Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14759-63
references.pmid: 9405686
references.citation: J Virol. 1997 Aug;71(8):5894-904
references.pmid: 9223479
references.citation: J Gen Virol. 2006 Jun;87(Pt 6):1465-1475
references.pmid: 16690911
references.citation: Proc Natl Acad Sci U S A. 1999 Aug 3;96(16):9281-6
references.pmid: 10430934
references.citation: Am J Physiol Lung Cell Mol Physiol. 2009 Jun;296(6):L936-46
references.pmid: 19304907
references.citation: PLoS One. 2012;7(1):e30152
references.pmid: 22253913
references.citation: Nat Protoc. 2010 Apr;5(4):725-38
references.pmid: 20360767
references.citation: J Virol. 1994 Mar;68(3):1667-74
references.pmid: 8107227
references.citation: J Gen Virol. 1990 Jun;71 ( Pt 6):1355-64
references.pmid: 2351958
references.citation: J Gen Virol. 2009 May;90(Pt 5):1202-1214
references.pmid: 19264603
references.citation: J Immunol. 2005 Apr 15;174(8):4972-8
references.pmid: 15814726
references.citation: PLoS One. 2012;7(10):e48451
references.pmid: 23119022
references.citation: BMC Bioinformatics. 2008 Jan 23;9:40
references.pmid: 18215316
references.citation: J Immunol. 2000 Jul 15;165(2):1074-81
references.pmid: 10878386
references.citation: Proc Natl Acad Sci U S A. 2009 Feb 24;106(8):2880-5
references.pmid: 19196955
references.citation: Cell Mol Life Sci. 2008 Jun;65(11):1653-68
references.pmid: 18351291
references.citation: Annu Rev Immunol. 2011;29:351-97
references.pmid: 21219186
references.citation: J Virol. 2007 May;81(9):4732-43
references.pmid: 17314170
references.citation: Nat Rev Mol Cell Biol. 2002 Oct;3(10):753-66
references.pmid: 12360192
references.citation: J Virol. 2004 Dec;78(23):13370-5
references.pmid: 15542690
references.citation: J Gen Virol. 2009 Jan;90(Pt 1):21-32
references.pmid: 19088269
references.citation: J Virol. 1989 Jan;63(1):403-10
references.pmid: 2535741
references.citation: Science. 2007 Feb 9;315(5813):843-8
references.pmid: 17289996
references.citation: J Virol. 2007 Dec;81(23):13082-91
references.pmid: 17898071
references.citation: Science. 2006 Jul 14;313(5784):187-91
references.pmid: 16840692
references.citation: J Biol Chem. 1991 Jul 5;266(19):12127-30
references.pmid: 1905715
references.citation: J Exp Med. 2008 Mar 17;205(3):669-84
references.pmid: 18332178
references.citation: J Virol. 2004 May;78(10):5103-12
references.pmid: 15113892
references.citation: Science. 2006 Jul 14;313(5784):217-20
references.pmid: 16840698
references.citation: PLoS One. 2008 Jul 30;3(7):e2811
references.pmid: 18665235