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J Virol. Mar 1997; 71(3): 2163–2170.
PMCID: PMC191323

Site-directed and linker insertion mutagenesis of herpes simplex virus type 1 glycoprotein H.

Abstract

The gH-gL complex of herpes simplex virus type 1 (HSV-1) is essential for virion infectivity and virus-induced cell fusion, but functional domains of the gH molecule remain to be defined. We have addressed this question by mutagenesis. A set of linker insertion mutants in HSV-1 gH was generated and tested in transient assays for their ability to complement a gH-negative virus. Insertions at three sites in the C-terminal third of the external domain affected the ability of gH to function in cell-cell fusion and virus entry, while insertions at six sites in the N-terminal half of the external domain induced conformational changes in gH such that it was not recognized by monoclonal antibody LP11, although expression at the cell surface was unchanged. A recombinant virus in which a potential integrin-binding motif, RGD, in gH was changed to the triplet RGE entered cells as efficiently as the wild type, indicating that HSV-1 entry is not mediated by means of the gH-RGD motif binding to cell surface integrins. Furthermore, mutagenesis of the glycosylation site which is positionally conserved in all herpesvirus gH sequences in close proximity to the transmembrane domain generated a recombinant virus that grew in vitro with wild-type single-step kinetics.

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Selected References

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  • Anderson RA, Liu DX, Gompels UA. Definition of a human herpesvirus-6 betaherpesvirus-specific domain in glycoprotein gH that governs interaction with glycoprotein gL: substitution of human cytomegalovirus glycoproteins permits group-specific complex formation. Virology. 1996 Mar 15;217(2):517–526. [PubMed]
  • Browne H, Baxter V, Minson T. Analysis of protective immune responses to the glycoprotein H-glycoprotein L complex of herpes simplex virus type 1. J Gen Virol. 1993 Dec;74(Pt 12):2813–2817. [PubMed]
  • Browne H, Bell S, Minson T, Wilson DW. An endoplasmic reticulum-retained herpes simplex virus glycoprotein H is absent from secreted virions: evidence for reenvelopment during egress. J Virol. 1996 Jul;70(7):4311–4316. [PMC free article] [PubMed]
  • Browne HM, Bruun BC, Minson AC. Characterization of herpes simplex virus type 1 recombinants with mutations in the cytoplasmic tail of glycoprotein H. J Gen Virol. 1996 Oct;77(Pt 10):2569–2573. [PubMed]
  • Cai WZ, Person S, DebRoy C, Gu BH. Functional regions and structural features of the gB glycoprotein of herpes simplex virus type 1. An analysis of linker insertion mutants. J Mol Biol. 1988 Jun 5;201(3):575–588. [PubMed]
  • Chen C, Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. [PMC free article] [PubMed]
  • Chiang HY, Cohen GH, Eisenberg RJ. Identification of functional regions of herpes simplex virus glycoprotein gD by using linker-insertion mutagenesis. J Virol. 1994 Apr;68(4):2529–2543. [PMC free article] [PubMed]
  • Davis-Poynter N, Bell S, Minson T, Browne H. Analysis of the contributions of herpes simplex virus type 1 membrane proteins to the induction of cell-cell fusion. J Virol. 1994 Nov;68(11):7586–7590. [PMC free article] [PubMed]
  • Desai PJ, Schaffer PA, Minson AC. Excretion of non-infectious virus particles lacking glycoprotein H by a temperature-sensitive mutant of herpes simplex virus type 1: evidence that gH is essential for virion infectivity. J Gen Virol. 1988 Jun;69(Pt 6):1147–1156. [PubMed]
  • Digard P, Bebrin WR, Weisshart K, Coen DM. The extreme C terminus of herpes simplex virus DNA polymerase is crucial for functional interaction with processivity factor UL42 and for viral replication. J Virol. 1993 Jan;67(1):398–406. [PMC free article] [PubMed]
  • D'Souza SE, Ginsberg MH, Plow EF. Arginyl-glycyl-aspartic acid (RGD): a cell adhesion motif. Trends Biochem Sci. 1991 Jul;16(7):246–250. [PubMed]
  • Forrester A, Farrell H, Wilkinson G, Kaye J, Davis-Poynter N, Minson T. Construction and properties of a mutant of herpes simplex virus type 1 with glycoprotein H coding sequences deleted. J Virol. 1992 Jan;66(1):341–348. [PMC free article] [PubMed]
  • Fox G, Parry NR, Barnett PV, McGinn B, Rowlands DJ, Brown F. The cell attachment site on foot-and-mouth disease virus includes the amino acid sequence RGD (arginine-glycine-aspartic acid). J Gen Virol. 1989 Mar;70(Pt 3):625–637. [PubMed]
  • Gompels UA, Carss AL, Saxby C, Hancock DC, Forrester A, Minson AC. Characterization and sequence analyses of antibody-selected antigenic variants of herpes simplex virus show a conformationally complex epitope on glycoprotein H. J Virol. 1991 May;65(5):2393–2401. [PMC free article] [PubMed]
  • Gompels UA, Craxton MA, Honess RW. Conservation of glycoprotein H (gH) in herpesviruses: nucleotide sequence of the gH gene from herpesvirus saimiri. J Gen Virol. 1988 Nov;69(Pt 11):2819–2829. [PubMed]
  • Gompels U, Minson A. The properties and sequence of glycoprotein H of herpes simplex virus type 1. Virology. 1986 Sep;153(2):230–247. [PubMed]
  • Hutchinson L, Browne H, Wargent V, Davis-Poynter N, Primorac S, Goldsmith K, Minson AC, Johnson DC. 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. 1992 Apr;66(4):2240–2250. [PMC free article] [PubMed]
  • Kaye JF, Gompels UA, Minson AC. Glycoprotein H of human cytomegalovirus (HCMV) forms a stable complex with the HCMV UL115 gene product. J Gen Virol. 1992 Oct;73(Pt 10):2693–2698. [PubMed]
  • Keller PM, Davison AJ, Lowe RS, Riemen MW, Ellis RW. Identification and sequence of the gene encoding gpIII, a major glycoprotein of varicella-zoster virus. Virology. 1987 Apr;157(2):526–533. [PubMed]
  • Klupp BG, Baumeister J, Karger A, Visser N, Mettenleiter TC. Identification and characterization of a novel structural glycoprotein in pseudorabies virus, gL. J Virol. 1994 Jun;68(6):3868–3878. [PMC free article] [PubMed]
  • Klupp BG, Mettenleiter TC. Sequence and expression of the glycoprotein gH gene of pseudorabies virus. Virology. 1991 Jun;182(2):732–741. [PubMed]
  • Kunkel TA. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. [PMC free article] [PubMed]
  • Liu DX, Cavanagh D, Green P, Inglis SC. A polycistronic mRNA specified by the coronavirus infectious bronchitis virus. Virology. 1991 Oct;184(2):531–544. [PubMed]
  • Liu DX, Gompels UA, Foa-Tomasi L, Campadelli-Fiume G. Human herpesvirus-6 glycoprotein H and L homologs are components of the gp100 complex and the gH external domain is the target for neutralizing monoclonal antibodies. Virology. 1993 Nov;197(1):12–22. [PubMed]
  • Liu DX, Gompels UA, Nicholas J, Lelliott C. Identification and expression of the human herpesvirus 6 glycoprotein H and interaction with an accessory 40K glycoprotein. J Gen Virol. 1993 Sep;74(Pt 9):1847–1857. [PubMed]
  • McGeoch DJ, Cook S, Dolan A, Jamieson FE, Telford EA. Molecular phylogeny and evolutionary timescale for the family of mammalian herpesviruses. J Mol Biol. 1995 Mar 31;247(3):443–458. [PubMed]
  • McGeoch DJ, Davison AJ. DNA sequence of the herpes simplex virus type 1 gene encoding glycoprotein gH, and identification of homologues in the genomes of varicella-zoster virus and Epstein-Barr virus. Nucleic Acids Res. 1986 May 27;14(10):4281–4292. [PMC free article] [PubMed]
  • Meyer AL, Petrovskis EA, Duffus WP, Thomsen DR, Post LE. Cloning and sequence of an infectious bovine rhinotracheitis virus (BHV-1) gene homologous to glycoprotein H of herpes simplex virus. Biochim Biophys Acta. 1991 Oct 8;1090(2):267–269. [PubMed]
  • Nicolson L, Cullinane AA, Onions DE. The nucleotide sequence of an equine herpesvirus 4 gene homologue of the herpes simplex virus 1 glycoprotein H gene. J Gen Virol. 1990 Aug;71(Pt 8):1793–1800. [PubMed]
  • Robertson GR, Scott NA, Miller JM, Sabine M, Zheng M, Bell CW, Whalley JM. Sequence characteristics of a gene in equine herpesvirus 1 homologous to glycoprotein H of herpes simplex virus. DNA Seq. 1991;1(4):241–249. [PubMed]
  • Roivainen M, Hyypiä T, Piirainen L, Kalkkinen N, Stanway G, Hovi T. RGD-dependent entry of coxsackievirus A9 into host cells and its bypass after cleavage of VP1 protein by intestinal proteases. J Virol. 1991 Sep;65(9):4735–4740. [PMC free article] [PubMed]
  • Shieh MT, Spear PG. Herpesvirus-induced cell fusion that is dependent on cell surface heparan sulfate or soluble heparin. J Virol. 1994 Feb;68(2):1224–1228. [PMC free article] [PubMed]
  • Wickham TJ, Mathias P, Cheresh DA, Nemerow GR. Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell. 1993 Apr 23;73(2):309–319. [PubMed]
  • Wilson DW, Davis-Poynter N, Minson AC. Mutations in the cytoplasmic tail of herpes simplex virus glycoprotein H suppress cell fusion by a syncytial strain. J Virol. 1994 Nov;68(11):6985–6993. [PMC free article] [PubMed]
  • WuDunn D, Spear PG. Initial interaction of herpes simplex virus with cells is binding to heparan sulfate. J Virol. 1989 Jan;63(1):52–58. [PMC free article] [PubMed]
  • Yaswen LR, Stephens EB, Davenport LC, Hutt-Fletcher LM. Epstein-Barr virus glycoprotein gp85 associates with the BKRF2 gene product and is incompletely processed as a recombinant protein. Virology. 1993 Aug;195(2):387–396. [PubMed]

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