title: Role of herpes simplex virus 1 immediate early protein ICP22 in viral nuclear egress.
abstract: In order to investigate the novel function(s) of the herpes simplex virus 1 (HSV-1) immediate early protein ICP22, we screened for ICP22-binding proteins in HSV-1-infected cells. Our results were as follows. (i) Tandem affinity purification of ICP22 from infected cells, coupled with mass spectrometry-based proteomics and subsequent analyses, demonstrates that ICP22 forms a complex(es) with the HSV-1 proteins UL31, UL34, UL47 (or VP13/14), and/or Us3. All these proteins were previously reported to be important for viral egress through the nuclear membrane. (ii) ICP22 colocalizes with UL31 and UL34 at the nuclear membrane in wild-type HSV-1-infected cells. (iii) The UL31-null mutation prevents the targeting of ICP22 to the nuclear membrane. (iv) The ICP22-null mutation resulted in UL31 and UL34 being mislocalized in the endoplasmic reticulum (in addition to the nuclear membrane) and significantly reduced numbers of primary enveloped virions in the perinuclear space, although capsids accumulated in the nuclei. Collectively, these results suggest that (i) ICP22 interacts with HSV-1 regulators of nuclear egress, including UL31, UL34, UL47, and Us3 in HSV-1-infected cells; (ii) UL31 mediates the recruitment and anchorage of ICP22 at the nuclear membrane; and (iii) ICP22 plays a regulatory role in HSV-1 primary envelopment, probably by interacting with and regulating UL31 and UL34. Here we report a previously unknown function for ICP22 in the regulation of HSV-1 nuclear egress.
  
  
  OBJECTIVE
  The herpes simplex virus 1 (HSV-1) immediate early protein ICP22 is recognized primarily as a regulator of viral gene expression. In this study, we show that ICP22 interacts with the HSV-1 proteins UL31 and UL34, which play crucial roles at the nuclear membrane in HSV-1 primary envelopment during viral nuclear egress. We also demonstrate that UL31 is required for the targeting of ICP22 to the nuclear membrane and that ICP22 is required for the correct localization of UL31 and/or UL34. Furthermore, we confirm that ICP22 is required for efficient HSV-1 primary envelopment during viral nuclear egress. Thus, we report, for the first time, that ICP22 plays a regulatory role in HSV-1 nuclear egress.
authors.affiliation: Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Yuhei
authors.initials: Y
authors.lastname: Maruzuru
authors.affiliation: Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Shindo
authors.affiliation: Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Zhuoming
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Medical Proteomics Laboratory, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Oyama
authors.affiliation: Medical Proteomics Laboratory, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Kozuka-Hata
authors.affiliation: Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Arii
authors.affiliation: Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Akihisa
authors.initials: A
authors.lastname: Kato
authors.affiliation: Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan ykawagu@ims.u-tokyo.ac.jp.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Kawaguchi
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism / Q000821:virology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000821:virology; D006561:Herpes Simplex / Q000378:metabolism* / Q000821:virology; D018259:Herpesvirus 1, Human / Q000502:physiology; D006801:Humans; D017874:Immediate-Early Proteins / Q000378:metabolism*; D008168:Lung / Q000166:cytology / Q000378:metabolism / Q000821:virology; D009685:Nuclear Envelope / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D021381:Protein Transport; D011817:Rabbits; D012867:Skin / Q000166:cytology / Q000378:metabolism / Q000821:virology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014709:Vero Cells; D014764:Viral Proteins / Q000378:metabolism*; D014771:Virion / Q000502:physiology; D019065:Virus Assembly / Q000502:physiology*
chemical_list: C057294:ICP22 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D009687:Nuclear Proteins; C108721:UL31 protein, Human herpesvirus 1; C403420:UL34 protein, Human herpesvirus 1; D014764:Viral Proteins; D017346:Protein Serine-Threonine Kinases; C095207:US3 protein, Human herpesvirus 1
references.citation: Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):11864-8
references.pmid: 7991547
references.citation: J Virol. 2011 Sep;85(18):9472-7
references.pmid: 21734043
references.citation: Virology. 2004 Nov 10;329(1):68-76
references.pmid: 15476875
references.citation: J Virol. 2008 Aug;82(16):8094-104
references.pmid: 18524819
references.citation: J Virol. 2008 Jan;82(2):617-29
references.pmid: 17959661
references.citation: J Virol. 1996 Jan;70(1):172-8
references.pmid: 8523523
references.citation: J Virol. 2008 Nov;82(21):10591-9
references.pmid: 18753202
references.citation: Proc Natl Acad Sci U S A. 2007 Apr 24;104(17):7241-6
references.pmid: 17426144
references.citation: J Virol. 2006 Jan;80(1):494-504
references.pmid: 16352573
references.citation: Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6701-5
references.pmid: 8393574
references.citation: J Virol. 2005 Mar;79(6):3797-806
references.pmid: 15731273
references.citation: J Virol. 1997 Feb;71(2):1133-9
references.pmid: 8995634
references.citation: J Virol. 2002 Sep;76(17):8939-52
references.pmid: 12163613
references.citation: J Virol. 1997 Feb;71(2):1019-24
references.pmid: 8995621
references.citation: J Virol. 1994 Feb;68(2):988-1001
references.pmid: 8289400
references.citation: Cell Microbiol. 2013 Feb;15(2):170-8
references.pmid: 23057731
references.citation: J Virol. 1999 May;73(5):4305-15
references.pmid: 10196329
references.citation: J Virol. 2009 Jun;83(11):5773-83
references.pmid: 19297494
references.citation: J Virol. 1997 Nov;71(11):8307-15
references.pmid: 9343183
references.citation: J Virol. 2014 Feb;88(4):2359-64
references.pmid: 24335305
references.citation: J Virol. 2011 Aug;85(15):7513-22
references.pmid: 21632758
references.citation: J Virol. 2014 May;88(9):4657-67
references.pmid: 24522907
references.citation: J Virol. 2011 Nov;85(22):11972-80
references.pmid: 21880766
references.citation: J Virol. 2004 Jan;78(1):399-412
references.pmid: 14671121
references.citation: J Virol. 1985 Aug;55(2):338-46
references.pmid: 2991560
references.citation: J Virol. 2010 Jan;84(1):153-62
references.pmid: 19846518
references.citation: J Virol. 1997 Oct;71(10):7328-36
references.pmid: 9311810
references.citation: J Virol. 2006 Oct;80(19):9381-90
references.pmid: 16973544
references.citation: J Virol. 2000 Jan;74(1):117-29
references.pmid: 10590098
references.citation: J Virol. 2012 Jan;86(1):594-8
references.pmid: 22013050
references.citation: J Virol. 2009 Nov;83(22):11624-34
references.pmid: 19740999
references.citation: J Virol. 1995 Sep;69(9):5550-9
references.pmid: 7637000
references.citation: J Virol. 2001 Sep;75(18):8803-17
references.pmid: 11507225
references.citation: J Virol. 2010 Mar;84(5):2384-94
references.pmid: 20032172
references.citation: Nat Rev Microbiol. 2011 May;9(5):382-94
references.pmid: 21494278
references.citation: J Virol. 2004 Jun;78(11):5564-75
references.pmid: 15140953
references.citation: J Virol. 2007 Jun;81(12):6459-70
references.pmid: 17428859
references.citation: J Virol. 2008 Jul;82(13):6172-89
references.pmid: 18417577
references.citation: J Virol. 1991 Nov;65(11):5757-64
references.pmid: 1656069
references.citation: Nature. 2010 Oct 14;467(7317):859-62
references.pmid: 20944748
references.citation: J Virol. 2013 Aug;87(16):9323-32
references.pmid: 23785201
references.citation: J Virol. 2003 Jan;77(2):1382-91
references.pmid: 12502854
references.citation: Virology. 2006 Apr 10;347(2):261-76
references.pmid: 16427676
references.citation: J Virol. 2005 Jul;79(14):9325-31
references.pmid: 15994828
references.citation: J Virol. 2005 Jun;79(11):6757-62
references.pmid: 15890914
references.citation: J Virol. 2008 Jun;82(11):5198-211
references.pmid: 18353954
references.citation: Virology. 1997 Aug 4;234(2):226-34
references.pmid: 9268153
references.citation: J Virol. 2011 Sep;85(18):9599-613
references.pmid: 21734045