Skip to main content
SpringerLink
Log in

Characterization of the subcellular localization of herpes simplex virus type 1 proteins in living cells

  • Rapid Communication
  • Published:
Medical Microbiology and Immunology Aims and scope Submit manuscript

Abstract

In this study, we presented the construction of a library of expression clones for the herpes simplex virus type 1 (HSV-1) proteome and subcellular localization map of HSV-1 proteins in living cells using yellow fluorescent protein (YFP) fusion proteins. As a result, 21 proteins showed cytoplasmic or subcytoplasmic localization, 16 proteins showed nuclear or subnuclear localization, and others were present both in the nucleus and cytoplasm. Interestingly, most capsid proteins showed enriched or exclusive localization in the nucleus, and most of the envelope proteins showed cytoplasmic localization, suggesting that subcellular localization of the proteins correlated with their functions during virus replication. These results present a subcellular localization map of HSV-1 proteins in living cells, which provide useful information to further characterize the functions of these proteins.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

References

  1. Loret S, Guay G, Lippe R (2008) Comprehensive characterization of extracellular herpes simplex virus type 1 virions. J Virol 82(17):8605–8618. doi:JVI.00904-08[pii]10.1128/JVI.00904-08

    Article  CAS  PubMed  Google Scholar 

  2. Simpson JC, Neubrand VE, Wiemann S, Pepperkok R (2001) Illuminating the human genome. Histochem Cell Biol 115(1):23–29

    CAS  PubMed  Google Scholar 

  3. Tanaka M, Kagawa H, Yamanashi Y, Sata T, Kawaguchi Y (2003) Construction of an excisable bacterial artificial chromosome containing a full-length infectious clone of herpes simplex virus type 1: viruses reconstituted from the clone exhibit wild-type properties in vitro and in vivo. J Virol 77(2):1382–1391

    Article  CAS  PubMed  Google Scholar 

  4. Barnard EC, Brown G, Stow ND (1997) Deletion mutants of the herpes simplex virus type 1 UL8 protein: effect on DNA synthesis and ability to interact with and influence the intracellular localization of the UL5 and UL52 proteins. Virology 237(1):97–106. doi:S0042-6822(97)98763-8[pii]10.1006/viro.1997.8763

    Article  CAS  PubMed  Google Scholar 

  5. Yu D, Weller SK (1998) Genetic analysis of the UL 15 gene locus for the putative terminase of herpes simplex virus type 1. Virology 243(1):32–44. doi:S0042-6822(98)99041-9[pii]10.1006/viro.1998.9041

    Article  CAS  PubMed  Google Scholar 

  6. Roberts AP, Abaitua F, O’Hare P, McNab D, Rixon FJ, Pasdeloup D (2009) Differing roles of inner tegument proteins PUl36 and PUl37 during entry of herpes simplex virus type 1. J Virol 83(1):105–116. doi:JVI.01032-08[pii]10.1128/JVI.01032-08

    Article  CAS  PubMed  Google Scholar 

  7. Boutell C, Everett RD (2003) The herpes simplex virus type 1 (HSV-1) regulatory protein ICP0 interacts with and ubiquitinates p53. J Biol Chem 278(38):36596–36602. doi:10.1074/jbc.M300776200M300776200[pii]

    Article  CAS  PubMed  Google Scholar 

  8. Xing J, Wu F, Pan W, Zheng C (2010) Molecular anatomy of subcellular localization of HSV-1 tegument protein US11 in living cells. Virus Res 153(1):71–81

    Article  CAS  PubMed  Google Scholar 

  9. Salsman J, Zimmerman N, Chen T, Domagala M, Frappier L (2008) Genome-wide screen of three herpesviruses for protein subcellular localization and alteration of PML nuclear bodies. PLoS Pathog 4(7):e1000100. doi:10.1371/journal.ppat.1000100

    Article  PubMed  Google Scholar 

  10. Calder JM, Stow EC, Stow ND (1992) On the cellular localization of the components of the herpes simplex virus type 1 helicase-primase complex and the viral origin-binding protein. J Gen Virol 73(Pt 3):531–538

    Article  CAS  PubMed  Google Scholar 

  11. Fraser KA, Rice SA (2007) Herpes simplex virus immediate-early protein ICP22 triggers loss of serine 2-phosphorylated RNA polymerase II. J Virol 81(10):5091–5101. doi:JVI.00184-07[pii]10.1128/JVI.00184-07

    Article  CAS  PubMed  Google Scholar 

  12. Burkham J, Coen DM, Hwang CB, Weller SK (2001) Interactions of herpes simplex virus type 1 with ND10 and recruitment of PML to replication compartments. J Virol 75(5):2353–2367. doi:10.1128/JVI.75.5.2353-2367.2001

    Article  CAS  PubMed  Google Scholar 

  13. Chen Y, Ciustea M, Ricciardi RP (2005) Processivity factor of KSHV contains a nuclear localization signal and binding domains for transporting viral DNA polymerase into the nucleus. Virology 340(2):183–191

    Article  CAS  PubMed  Google Scholar 

  14. Carter KL, Ward PL, Roizman B (1996) Characterization of the products of the U(L)43 gene of herpes simplex virus 1: potential implications for regulation of gene expression by antisense transcription. J Virol 70(11):7663–7668

    CAS  PubMed  Google Scholar 

  15. Samaniego LA, Neiderhiser L, DeLuca NA (1998) Persistence and expression of the herpes simplex virus genome in the absence of immediate-early proteins. J Virol 72(4):3307–3320

    CAS  PubMed  Google Scholar 

  16. Markovitz NS, Roizman B (2000) Small dense nuclear bodies are the site of localization of herpes simplex virus 1 U(L)3 and U(L)4 proteins and of ICP22 only when the latter protein is present. J Virol 74(1):523–528

    Article  CAS  PubMed  Google Scholar 

  17. Reynolds AE, Ryckman BJ, Baines JD, Zhou Y, Liang L, Roller RJ (2001) U(L)31 and U(L)34 proteins of herpes simplex virus type 1 form a complex that accumulates at the nuclear rim and is required for envelopment of nucleocapsids. J Virol 75(18):8803–8817

    Article  CAS  PubMed  Google Scholar 

  18. Vittone V, Diefenbach E, Triffett D, Douglas MW, Cunningham AL, Diefenbach RJ (2005) Determination of interactions between tegument proteins of herpes simplex virus type 1. J Virol 79(15):9566–9571. doi:79/15/9566[pii]10.1128/JVI.79.15.9566-9571.2005

    Article  CAS  PubMed  Google Scholar 

  19. Roizman B (1999) Hsv gene functions: what have we learned that could be generally applicable to its near and distant cousins? Acta Virol 43(2–3):75–80

    CAS  PubMed  Google Scholar 

  20. Everett RD, Sourvinos G, Orr A (2003) Recruitment of herpes simplex virus type 1 transcriptional regulatory protein ICP4 into foci juxtaposed to ND10 in live, infected cells. J Virol 77(6):3680–3689

    Article  CAS  PubMed  Google Scholar 

  21. Harland J, Dunn P, Cameron E, Conner J, Brown SM (2003) The herpes simplex virus (HSV) protein ICP34.5 is a virion component that forms a DNA-binding complex with proliferating cell nuclear antigen and HSV replication proteins. J Neurovirol 9(4):477–488. doi:AXPGX9NE3C7F07HM[pii]

    CAS  PubMed  Google Scholar 

  22. Hutchinson L, Browne H, Wargent V, Davis-Poynter N, Primorac S, Goldsmith K, Minson AC, Johnson DC (1992) A novel herpes simplex virus glycoprotein, gL, forms a complex with glycoprotein H (gH) and affects normal folding and surface expression of gH. J Virol 66(4):2240–2250

    CAS  PubMed  Google Scholar 

  23. Winters TA, Williams MV (1993) Purification and characterization of the herpes simplex virus type 2-encoded uracil-DNA glycosylase. Virology 195(2):315–326. doi:S0042-6822(83)71382-6[pii]10.1006/viro.1993.1382

    Article  CAS  PubMed  Google Scholar 

  24. Yamada H, Jiang YM, Oshima S, Wada K, Goshima F, Daikoku T, Nishiyama Y (1998) Characterization of the UL4 gene product of herpes simplex virus type 2. Arch Virol 143(6):1199–1207

    Article  CAS  PubMed  Google Scholar 

  25. White CA, Stow ND, Patel AH, Hughes M, Preston VG (2003) Herpes simplex virus type 1 portal protein UL6 interacts with the putative terminase subunits UL15 and UL28. J Virol 77(11):6351–6358

    Article  CAS  PubMed  Google Scholar 

  26. Tanaka M, Sata T, Kawaguchi Y (2008) The product of the herpes simplex virus 1 UL7 gene interacts with a mitochondrial protein, adenine nucleotide translocator 2. Virol J 5:125. doi:1743-422X-5-125[pii]10.1186/1743-422X-5-125

    Article  PubMed  Google Scholar 

  27. Link MA, Silva LA, Schaffer PA (2007) Cathepsin B mediates cleavage of herpes simplex virus type 1 origin binding protein (OBP) to yield OBPC-1, and cleavage is dependent upon viral DNA replication. J Virol 81(17):9175–9182. doi:JVI.00676-07[pii]10.1128/JVI.00676-07

    Article  CAS  PubMed  Google Scholar 

  28. Malik AK, Martinez R, Muncy L, Carmichael EP, Weller SK (1992) Genetic analysis of the herpes simplex virus type 1 UL9 gene: isolation of a LacZ insertion mutant and expression in eukaryotic cells. Virology 190(2):702–715

    Article  CAS  PubMed  Google Scholar 

  29. Crump CM, Bruun B, Bell S, Pomeranz LE, Minson T, Browne HM (2004) Alphaherpesvirus glycoprotein M causes the relocalization of plasma membrane proteins. J Gen Virol 85(Pt 12):3517–3527. doi:85/12/3517[pii]10.1099/vir.0.80361-0

    Article  CAS  PubMed  Google Scholar 

  30. Zhang J, Nagel CH, Sodeik B, Lippe R (2009) Early, active, and specific localization of herpes simplex virus type 1 gM to nuclear membranes. J Virol 83(24):12984–12997. doi:JVI.01180-09[pii]10.1128/JVI.01180-09

    Article  CAS  PubMed  Google Scholar 

  31. Fulmer PA, Melancon JM, Baines JD, Kousoulas KG (2007) UL20 protein functions precede and are required for the UL11 functions of herpes simplex virus type 1 cytoplasmic virion envelopment. J Virol 81(7):3097–3108. doi:JVI.02201-06[pii]10.1128/JVI.02201-06

    Article  CAS  PubMed  Google Scholar 

  32. Corcoran JA, Saffran HA, Duguay BA, Smiley JR (2009) Herpes simplex virus UL12.5 targets mitochondria through a mitochondrial localization sequence proximal to the N terminus. J Virol 83(6):2601–2610. doi:JVI.02087-08[pii]10.1128/JVI.02087-08

    Article  CAS  PubMed  Google Scholar 

  33. Yamauchi Y, Kiriyama K, Kubota N, Kimura H, Usukura J, Nishiyama Y (2008) The UL14 tegument protein of herpes simplex virus type 1 is required for efficient nuclear transport of the alpha transinducing factor VP16 and viral capsids. J Virol 82(3):1094–1106. doi:JVI.01226-07[pii]10.1128/JVI.01226-07

    Article  CAS  PubMed  Google Scholar 

  34. Meckes DG Jr, Wills JW (2007) Dynamic interactions of the UL16 tegument protein with the capsid of herpes simplex virus. J Virol 81(23):13028–13036. doi:JVI.01306-07[pii]10.1128/JVI.01306-07

    Article  CAS  PubMed  Google Scholar 

  35. Taus NS, Salmon B, Baines JD (1998) The herpes simplex virus 1 UL 17 gene is required for localization of capsids and major and minor capsid proteins to intranuclear sites where viral DNA is cleaved and packaged. Virology 252(1):115–125. doi:S0042-6822(98)99439-9[pii]10.1006/viro.1998.9439

    Article  CAS  PubMed  Google Scholar 

  36. Rixon FJ, Addison C, McGregor A, Macnab SJ, Nicholson P, Preston VG, Tatman JD (1996) Multiple interactions control the intracellular localization of the herpes simplex virus type 1 capsid proteins. J Gen Virol 77(Pt 9):2251–2260

    Article  CAS  PubMed  Google Scholar 

  37. Foster TP, Chouljenko VN, Kousoulas KG (2008) Functional and physical interactions of the herpes simplex virus type 1 UL20 membrane protein with glycoprotein K. J Virol 82(13):6310–6323. doi:JVI.00147-08[pii]10.1128/JVI.00147-08

    Article  CAS  PubMed  Google Scholar 

  38. Takakuwa H, Goshima F, Koshizuka T, Murata T, Daikoku T, Nishiyama Y (2001) Herpes simplex virus encodes a virion-associated protein which promotes long cellular processes in over-expressing cells. Genes Cells 6(11):955–966. doi:475[pii]

    Article  CAS  PubMed  Google Scholar 

  39. Gianni T, Martelli PL, Casadio R, Campadelli-Fiume G (2005) The ectodomain of herpes simplex virus glycoprotein H contains a membrane alpha-helix with attributes of an internal fusion peptide, positionally conserved in the herpesviridae family. J Virol 79(5):2931–2940. doi:79/5/2931[pii]10.1128/JVI.79.5.2931-2940.2005

    Article  CAS  PubMed  Google Scholar 

  40. Degreve B, Johansson M, De Clercq E, Karlsson A, Balzarini J (1998) Differential intracellular compartmentalization of herpetic thymidine kinases (TKs) in TK gene-transfected tumor cells: molecular characterization of the nuclear localization signal of herpes simplex virus type 1 TK. J Virol 72(12):9535–9543

    CAS  PubMed  Google Scholar 

  41. Lymberopoulos MH, Pearson A (2007) Involvement of UL24 in herpes-simplex-virus-1-induced dispersal of nucleolin. Virology 363(2):397–409. doi:S0042-6822(07)00043-8[pii]10.1016/j.virol.2007.01.028

    Article  CAS  PubMed  Google Scholar 

  42. Ogasawara M, Suzutani T, Yoshida I, Azuma M (2001) Role of the UL25 gene product in packaging DNA into the herpes simplex virus capsid: location of UL25 product in the capsid and demonstration that it binds DNA. J Virol 75(3):1427–1436. doi:10.1128/JVI.75.3.1427-1436.2001

    Article  CAS  PubMed  Google Scholar 

  43. Goshima F, Watanabe D, Takakuwa H, Wada K, Daikoku T, Yamada M, Nishiyama Y (2000) Herpes simplex virus UL17 protein is associated with B capsids and colocalizes with ICP35 and VP5 in infected cells. Arch Virol 145(2):417–426

    Article  CAS  PubMed  Google Scholar 

  44. Beitia Ortiz de Zarate I, Kaelin K, Rozenberg F (2004) Effects of mutations in the cytoplasmic domain of herpes simplex virus type 1 glycoprotein B on intracellular transport and infectivity. J Virol 78(3):1540–1551

    Article  PubMed  Google Scholar 

  45. Beard PM, Taus NS, Baines JD (2002) DNA cleavage and packaging proteins encoded by genes U(L)28, U(L)15, and U(L)33 of herpes simplex virus type 1 form a complex in infected cells. J Virol 76(10):4785–4791

    Article  CAS  PubMed  Google Scholar 

  46. Malik AK, Shao L, Shanley JD, Weller SK (1996) Intracellular localization of the herpes simplex virus type-1 origin binding protein, UL9. Virology 224(2):380–389. doi:S0042-6822(96)90545-0[pii]10.1006/viro.1996.0545

    Article  CAS  PubMed  Google Scholar 

  47. Chang YE, Poon AP, Roizman B (1996) Properties of the protein encoded by the UL32 open reading frame of herpes simplex virus 1. J Virol 70(6):3938–3946

    CAS  PubMed  Google Scholar 

  48. Kato A, Yamamoto M, Ohno T, Tanaka M, Sata T, Nishiyama Y, Kawaguchi Y (2006) Herpes simplex virus 1-encoded protein kinase UL13 phosphorylates viral US3 protein kinase and regulates nuclear localization of viral envelopment factors UL34 and UL31. J Virol 80(3):1476–1486. doi:80/3/1476[pii]10.1128/JVI.80.3.1476-1486.2006

    Article  CAS  PubMed  Google Scholar 

  49. Yedowitz JC, Kotsakis A, Schlegel EF, Blaho JA (2005) Nuclear localizations of the herpes simplex virus type 1 tegument proteins VP13/14, vhs, and VP16 precede VP22-dependent microtubule reorganization and VP22 nuclear import. J Virol 79(8):4730–4743. doi:79/8/4730[pii]10.1128/JVI.79.8.4730-4743.2005

    Article  CAS  PubMed  Google Scholar 

  50. Olivo PD, Nelson NJ, Challberg MD (1989) Herpes simplex virus type 1 gene products required for DNA replication: identification and overexpression. J Virol 63(1):196–204

    CAS  PubMed  Google Scholar 

  51. Ward PL, Barker DE, Roizman B (1996) A novel herpes simplex virus 1 gene, UL43.5, maps antisense to the UL43 gene and encodes a protein which colocalizes in nuclear structures with capsid proteins. J Virol 70(5):2684–2690

    CAS  PubMed  Google Scholar 

  52. Murphy MA, Bucks MA, O’Regan KJ, Courtney RJ (2008) The HSV-1 tegument protein pUL46 associates with cellular membranes and viral capsids. Virology 376(2):279–289. doi:S0042-6822(08)00190-6[pii]10.1016/j.virol.2008.03.018

    Article  CAS  PubMed  Google Scholar 

  53. Donnelly M, Elliott G (2001) Nuclear localization and shuttling of herpes simplex virus tegument protein VP13/14. J Virol 75(6):2566–2574. doi:10.1128/JVI.75.6.2566-2574.2001

    Article  CAS  PubMed  Google Scholar 

  54. Morrison EE, Stevenson AJ, Wang YF, Meredith DM (1998) Differences in the intracellular localization and fate of herpes simplex virus tegument proteins early in the infection of vero cells. J Gen Virol 79(Pt 10):2517–2528

    CAS  PubMed  Google Scholar 

  55. Blouin A, Blaho JA (2001) Assessment of the subcellular localization of the herpes simplex virus structural protein VP22 in the absence of other viral gene products. Virus Res 81(1–2):57–68. doi:S0168170201003550[pii]

    Article  CAS  PubMed  Google Scholar 

  56. Nozawa N, Daikoku T, Koshizuka T, Yamauchi Y, Yoshikawa T, Nishiyama Y (2003) Subcellular localization of herpes simplex virus type 1 UL51 protein and role of palmitoylation in Golgi apparatus targeting. J Virol 77(5):3204–3216

    Article  CAS  PubMed  Google Scholar 

  57. Hutchinson L, Roop-Beauchamp C, Johnson DC (1995) Herpes simplex virus glycoprotein K is known to influence fusion of infected cells, yet is not on the cell surface. J Virol 69(7):4556–4563

    CAS  PubMed  Google Scholar 

  58. Mears WE, Lam V, Rice SA (1995) Identification of nuclear and nucleolar localization signals in the herpes simplex virus regulatory protein ICP27. J Virol 69(2):935–947

    CAS  PubMed  Google Scholar 

  59. Yamada H, Jiang YM, Oshima S, Daikoku T, Yamashita Y, Tsurumi T, Nishiyama Y (1998) Characterization of the UL55 gene product of herpes simplex virus type 2. J Gen Virol 79(Pt 8):1989–1995

    CAS  PubMed  Google Scholar 

  60. Koshizuka T, Goshima F, Takakuwa H, Nozawa N, Daikoku T, Koiwai O, Nishiyama Y (2002) Identification and characterization of the UL56 gene product of herpes simplex virus type 2. J Virol 76(13):6718–6728

    Article  CAS  PubMed  Google Scholar 

  61. Cun W, Hong M, Liu LD, Dong CH, Luo J, Li QH (2006) Structural and functional characterization of herpes simplex virus 1 immediate-early protein infected-cell protein 22. J Biochem 140(1):67–73. doi:140/1/67[pii]10.1093/jb/mvj135

    Article  PubMed  Google Scholar 

  62. Goshima F, Watanabe D, Suzuki H, Takakuwa H, Yamada H, Nishiyama Y (2001) The US2 gene product of herpes simplex virus type 2 interacts with cytokeratin 18. Arch Virol 146(11):2201–2209

    Article  CAS  PubMed  Google Scholar 

  63. Ghiasi H, Nesburn AB, Cai S, Wechsler SL (1998) The US5 open reading frame of herpes simplex virus type 1 does encode a glycoprotein (gJ). Intervirology 41(2–3):91–97. doi:int41091[pii]

    Article  CAS  PubMed  Google Scholar 

  64. Whiteley A, Bruun B, Minson T, Browne H (1999) Effects of targeting herpes simplex virus type 1 gD to the endoplasmic reticulum and trans-Golgi network. J Virol 73(11):9515–9520

    CAS  PubMed  Google Scholar 

  65. Georgopoulou U, Kakkanas A, Miriagou V, Michaelidou A, Mavromara P (1995) Characterization of the US8.5 protein of herpes simplex virus. Arch Virol 140(12):2227–2241

    Article  CAS  PubMed  Google Scholar 

  66. Georgopoulou U, Michaelidou A, Roizman B, Mavromara-Nazos P (1993) Identification of a new transcriptional unit that yields a gene product within the unique sequences of the short component of the herpes simplex virus 1 genome. J Virol 67(7):3961–3968

    CAS  PubMed  Google Scholar 

  67. Diaz-Latoud C, Diaz JJ, Fabre-Jonca N, Kindbeiter K, Madjar JJ, Arrigo AP (1997) Herpes simplex virus US11 protein enhances recovery of protein synthesis and survival in heat shock treated HeLa cells. Cell Stress Chaperones 2(2):119–131

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by the Major State Basic Research Development Program of China (2010CB530105 and 2011CB504802); the National Natural Science Foundation of China (30900059, 30870120 and 81000736); the Start-up Fund of the Hundred Talents Program of the Chinese Academy of Sciences (20071010-141).

Author information

Authors and Affiliations

  1. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 430071, Wuhan, China

    Junji Xing, Shuai Wang, You Li, Hong Guo, Lei Zhao, Weiwei Pan, Fusen Lin, Huifang Zhu, Lei Wang, Meili Li, Lin Wang & Chunfu Zheng

  2. Wuhan Institute of Virology, Chinese Academy of Sciences, 44 Xiaohongshan, 430071, Wuchang, Wuhan, China

    Chunfu Zheng

Authors
  1. Junji Xing
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shuai Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. You Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hong Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lei Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Weiwei Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Fusen Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Huifang Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Lei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Meili Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Lin Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Chunfu Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chunfu Zheng.

Additional information

Junji Xing, Shuai Wang, You Li contributed equally to this study.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xing, J., Wang, S., Li, Y. et al. Characterization of the subcellular localization of herpes simplex virus type 1 proteins in living cells. Med Microbiol Immunol 200, 61–68 (2011). https://doi.org/10.1007/s00430-010-0175-9

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00430-010-0175-9

Keywords

Search

Navigation