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. 1995 Jun;69(6):3482–3489. doi: 10.1128/jvi.69.6.3482-3489.1995

Cell surface proteoglycans are not essential for infection by pseudorabies virus.

A Karger 1, A Saalmüller 1, F Tufaro 1, B W Banfield 1, T C Mettenleiter 1
PMCID: PMC189061  PMID: 7745695

Abstract

Cell surface proteoglycans, in particular those carrying heparan sulfate glycosaminoglycans, play a major role in primary attachment of herpesviruses to target cells. In pseudorabies virus (PrV), glycoprotein gC has been shown to represent the major heparan sulfate-binding virion envelope protein (T. C. Mettenleiter, L. Zsak, F. Zuckermann, N. Sugg, H. Kern, and T. Ben-Porat, J. Virol. 64:278-286, 1990). Since PrV gC is nonessential for viral infectivity in vitro and in vivo, either the interaction between virion envelope and cellular heparan sulfate is not necessary to mediate infection or other virion envelope proteins can substitute as heparan sulfate-binding components in the absence of gC. To answer these questions, we analyzed the infectivity of isogenic gC+ and gC- PrV on mouse L-cell derivatives with defects in glycosaminoglycan biosynthesis, using a rapid and sensitive fluorescence-based beta-galactosidase assay and single-cell counting in a fluorescence-activated cell sorter. Our data show that (i) in the virion, glycoprotein gC represents the only proteoglycan-binding envelope protein, and (ii) cellular proteoglycans are not essential for infectivity of PrV. Attachment studies using radiolabeled virions lacking either gC or the essential gD confirmed these results and demonstrated that PrV gD mainly contributes to binding of Pr virions to cell surface components other than proteoglycans. These data demonstrate the presence of a proteoglycan-independent mode of attachment for Pr virions leading to infectious entry into target cells.

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Selected References

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  1. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cai W. H., Gu B., Person S. Role of glycoprotein B of herpes simplex virus type 1 in viral entry and cell fusion. J Virol. 1988 Aug;62(8):2596–2604. doi: 10.1128/jvi.62.8.2596-2604.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Callebaut C., Krust B., Jacotot E., Hovanessian A. G. T cell activation antigen, CD26, as a cofactor for entry of HIV in CD4+ cells. Science. 1993 Dec 24;262(5142):2045–2050. doi: 10.1126/science.7903479. [DOI] [PubMed] [Google Scholar]
  4. Campadelli-Fiume G., Stirpe D., Boscaro A., Avitabile E., Foá-Tomasi L., Barker D., Roizman B. Glycoprotein C-dependent attachment of herpes simplex virus to susceptible cells leading to productive infection. Virology. 1990 Sep;178(1):213–222. doi: 10.1016/0042-6822(90)90396-9. [DOI] [PubMed] [Google Scholar]
  5. Compton T., Nowlin D. M., Cooper N. R. Initiation of human cytomegalovirus infection requires initial interaction with cell surface heparan sulfate. Virology. 1993 Apr;193(2):834–841. doi: 10.1006/viro.1993.1192. [DOI] [PubMed] [Google Scholar]
  6. Flynn S. J., Burgett B. L., Stein D. S., Wilkinson K. S., Ryan P. The amino-terminal one-third of pseudorabies virus glycoprotein gIII contains a functional attachment domain, but this domain is not required for the efficient penetration of Vero cells. J Virol. 1993 May;67(5):2646–2654. doi: 10.1128/jvi.67.5.2646-2654.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fuller A. O., Lee W. C. Herpes simplex virus type 1 entry through a cascade of virus-cell interactions requires different roles of gD and gH in penetration. J Virol. 1992 Aug;66(8):5002–5012. doi: 10.1128/jvi.66.8.5002-5012.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gruenheid S., Gatzke L., Meadows H., Tufaro F. Herpes simplex virus infection and propagation in a mouse L cell mutant lacking heparan sulfate proteoglycans. J Virol. 1993 Jan;67(1):93–100. doi: 10.1128/jvi.67.1.93-100.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hanham C. A., Zhao F., Tignor G. H. Evidence from the anti-idiotypic network that the acetylcholine receptor is a rabies virus receptor. J Virol. 1993 Jan;67(1):530–542. doi: 10.1128/jvi.67.1.530-542.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hanssens F. P., Nauwynck H. J., Pensaert M. B. Involvement of membrane-bound viral glycoproteins in adhesion of pseudorabies virus-infected cells. J Virol. 1993 Aug;67(8):4492–4496. doi: 10.1128/jvi.67.8.4492-4496.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Harouse J. M., Bhat S., Spitalnik S. L., Laughlin M., Stefano K., Silberberg D. H., Gonzalez-Scarano F. Inhibition of entry of HIV-1 in neural cell lines by antibodies against galactosyl ceramide. Science. 1991 Jul 19;253(5017):320–323. doi: 10.1126/science.1857969. [DOI] [PubMed] [Google Scholar]
  12. Herold B. C., Visalli R. J., Susmarski N., Brandt C. R., Spear P. G. Glycoprotein C-independent binding of herpes simplex virus to cells requires cell surface heparan sulphate and glycoprotein B. J Gen Virol. 1994 Jun;75(Pt 6):1211–1222. doi: 10.1099/0022-1317-75-6-1211. [DOI] [PubMed] [Google Scholar]
  13. Herold B. C., WuDunn D., Soltys N., Spear P. G. Glycoprotein C of herpes simplex virus type 1 plays a principal role in the adsorption of virus to cells and in infectivity. J Virol. 1991 Mar;65(3):1090–1098. doi: 10.1128/jvi.65.3.1090-1098.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hök M., Kjellén L., Johansson S. Cell-surface glycosaminoglycans. Annu Rev Biochem. 1984;53:847–869. doi: 10.1146/annurev.bi.53.070184.004215. [DOI] [PubMed] [Google Scholar]
  15. Johnson D. C., Burke R. L., Gregory T. Soluble forms of herpes simplex virus glycoprotein D bind to a limited number of cell surface receptors and inhibit virus entry into cells. J Virol. 1990 Jun;64(6):2569–2576. doi: 10.1128/jvi.64.6.2569-2576.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Johnson D. C., McDermott M. R., Chrisp C., Glorioso J. C. Pathogenicity in mice of herpes simplex virus type 2 mutants unable to express glycoprotein C. J Virol. 1986 Apr;58(1):36–42. doi: 10.1128/jvi.58.1.36-42.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. KAPLAN A. S., VATTER A. E. A comparison of herpes simplex and pseudorabies viruses. Virology. 1959 Apr;7(4):394–407. doi: 10.1016/0042-6822(59)90068-6. [DOI] [PubMed] [Google Scholar]
  18. Kaner R. J., Baird A., Mansukhani A., Basilico C., Summers B. D., Florkiewicz R. Z., Hajjar D. P. Fibroblast growth factor receptor is a portal of cellular entry for herpes simplex virus type 1. Science. 1990 Jun 15;248(4961):1410–1413. doi: 10.1126/science.2162560. [DOI] [PubMed] [Google Scholar]
  19. Karger A., Mettenleiter T. C. Glycoproteins gIII and gp50 play dominant roles in the biphasic attachment of pseudorabies virus. Virology. 1993 Jun;194(2):654–664. doi: 10.1006/viro.1993.1305. [DOI] [PubMed] [Google Scholar]
  20. Karger A., Saalmüller A., Tufaro F., Banfield B. W., Mettenleiter T. C. Cell surface proteoglycans are not essential for infection by pseudorabies virus. J Virol. 1995 Jun;69(6):3482–3489. doi: 10.1128/jvi.69.6.3482-3489.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kari B., Gehrz R. A human cytomegalovirus glycoprotein complex designated gC-II is a major heparin-binding component of the envelope. J Virol. 1992 Mar;66(3):1761–1764. doi: 10.1128/jvi.66.3.1761-1764.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Keay S., Merigan T. C., Rasmussen L. Identification of cell surface receptors for the 86-kilodalton glycoprotein of human cytomegalovirus. Proc Natl Acad Sci U S A. 1989 Dec;86(24):10100–10103. doi: 10.1073/pnas.86.24.10100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kimman T. G., de Wind N., Oei-Lie N., Pol J. M., Berns A. J., Gielkens A. L. Contribution of single genes within the unique short region of Aujeszky's disease virus (suid herpesvirus type 1) to virulence, pathogenesis and immunogenicity. J Gen Virol. 1992 Feb;73(Pt 2):243–251. doi: 10.1099/0022-1317-73-2-243. [DOI] [PubMed] [Google Scholar]
  24. Kritas S. K., Pensaert M. B., Mettenleiter T. C. Role of envelope glycoproteins gI, gp63 and gIII in the invasion and spread of Aujeszky's disease virus in the olfactory nervous pathway of the pig. J Gen Virol. 1994 Sep;75(Pt 9):2319–2327. doi: 10.1099/0022-1317-75-9-2319. [DOI] [PubMed] [Google Scholar]
  25. Langeland N., Moore L. J. Reduction of HSV-1 binding to BHK cells after treatment with phosphatidylinositol-specific phospholipase C. FEBS Lett. 1990 Dec 17;277(1-2):253–256. doi: 10.1016/0014-5793(90)80859-h. [DOI] [PubMed] [Google Scholar]
  26. Liang X. P., Babiuk L. A., Zamb T. J. Pseudorabies virus gIII and bovine herpesvirus 1 gIII share complementary functions. J Virol. 1991 Oct;65(10):5553–5557. doi: 10.1128/jvi.65.10.5553-5557.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Liang X., Babiuk L. A., Zamb T. J. Mapping of heparin-binding structures on bovine herpesvirus 1 and pseudorabies virus gIII glycoproteins. Virology. 1993 May;194(1):233–243. doi: 10.1006/viro.1993.1254. [DOI] [PubMed] [Google Scholar]
  28. Long D., Berson J. F., Cook D. G., Doms R. W. Characterization of human immunodeficiency virus type 1 gp120 binding to liposomes containing galactosylceramide. J Virol. 1994 Sep;68(9):5890–5898. doi: 10.1128/jvi.68.9.5890-5898.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lycke E., Johansson M., Svennerholm B., Lindahl U. Binding of herpes simplex virus to cellular heparan sulphate, an initial step in the adsorption process. J Gen Virol. 1991 May;72(Pt 5):1131–1137. doi: 10.1099/0022-1317-72-5-1131. [DOI] [PubMed] [Google Scholar]
  30. Marsh M., Helenius A. Virus entry into animal cells. Adv Virus Res. 1989;36:107–151. doi: 10.1016/S0065-3527(08)60583-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. McClain D. S., Fuller A. O. Cell-specific kinetics and efficiency of herpes simplex virus type 1 entry are determined by two distinct phases of attachment. Virology. 1994 Feb;198(2):690–702. doi: 10.1006/viro.1994.1081. [DOI] [PubMed] [Google Scholar]
  32. Mettenleiter T. C. Initiation and spread of alpha-herpesvirus infections. Trends Microbiol. 1994 Jan;2(1):2–4. doi: 10.1016/0966-842x(94)90335-2. [DOI] [PubMed] [Google Scholar]
  33. Mettenleiter T. C., Kern H., Rauh I. Isolation of a viable herpesvirus (pseudorabies virus) mutant specifically lacking all four known nonessential glycoproteins. Virology. 1990 Nov;179(1):498–503. doi: 10.1016/0042-6822(90)90324-k. [DOI] [PubMed] [Google Scholar]
  34. Mettenleiter T. C. Pseudorabies (Aujeszky's disease) virus: state of the art. August 1993. Acta Vet Hung. 1994;42(2-3):153–177. [PubMed] [Google Scholar]
  35. Mettenleiter T. C., Rauh I. A glycoprotein gX-beta-galactosidase fusion gene as insertional marker for rapid identification of pseudorabies virus mutants. J Virol Methods. 1990 Oct;30(1):55–65. doi: 10.1016/0166-0934(90)90043-f. [DOI] [PubMed] [Google Scholar]
  36. Mettenleiter T. C., Spear P. G. Glycoprotein gB (gII) of pseudorabies virus can functionally substitute for glycoprotein gB in herpes simplex virus type 1. J Virol. 1994 Jan;68(1):500–504. doi: 10.1128/jvi.68.1.500-504.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Mettenleiter T. C., Zsak L., Zuckermann F., Sugg N., Kern H., Ben-Porat T. Interaction of glycoprotein gIII with a cellular heparinlike substance mediates adsorption of pseudorabies virus. J Virol. 1990 Jan;64(1):278–286. doi: 10.1128/jvi.64.1.278-286.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Mirda D. P., Navarro D., Paz P., Lee P. L., Pereira L., Williams L. T. The fibroblast growth factor receptor is not required for herpes simplex virus type 1 infection. J Virol. 1992 Jan;66(1):448–457. doi: 10.1128/jvi.66.1.448-457.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Muggeridge M. I., Cohen G. H., Eisenberg R. J. Herpes simplex virus infection can occur without involvement of the fibroblast growth factor receptor. J Virol. 1992 Feb;66(2):824–830. doi: 10.1128/jvi.66.2.824-830.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Okazaki K., Matsuzaki T., Sugahara Y., Okada J., Hasebe M., Iwamura Y., Ohnishi M., Kanno T., Shimizu M., Honda E. BHV-1 adsorption is mediated by the interaction of glycoprotein gIII with heparinlike moiety on the cell surface. Virology. 1991 Apr;181(2):666–670. doi: 10.1016/0042-6822(91)90900-v. [DOI] [PubMed] [Google Scholar]
  41. Peeters B., de Wind N., Hooisma M., Wagenaar F., Gielkens A., Moormann R. Pseudorabies virus envelope glycoproteins gp50 and gII are essential for virus penetration, but only gII is involved in membrane fusion. J Virol. 1992 Feb;66(2):894–905. doi: 10.1128/jvi.66.2.894-905.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Powers L., Wilkinson K. S., Ryan P. Characterization of the prv43 gene of pseudorabies virus and demonstration that it is not required for virus growth in cell culture. Virology. 1994 Feb 15;199(1):81–88. doi: 10.1006/viro.1994.1099. [DOI] [PubMed] [Google Scholar]
  43. Rauh I., Mettenleiter T. C. Pseudorabies virus glycoproteins gII and gp50 are essential for virus penetration. J Virol. 1991 Oct;65(10):5348–5356. doi: 10.1128/jvi.65.10.5348-5356.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Saalmüller A., Mettenleiter T. C. Rapid identification and quantitation of cells infected by recombinant herpesvirus (pseudorabies virus) using a fluorescence-based beta-galactosidase assay and flow cytometry. J Virol Methods. 1993 Sep;44(1):99–108. doi: 10.1016/0166-0934(93)90012-g. [DOI] [PubMed] [Google Scholar]
  45. Sawitzky D., Hampl H., Habermehl K. O. Comparison of heparin-sensitive attachment of pseudorabies virus (PRV) and herpes simplex virus type 1 and identification of heparin-binding PRV glycoproteins. J Gen Virol. 1990 May;71(Pt 5):1221–1225. doi: 10.1099/0022-1317-71-5-1221. [DOI] [PubMed] [Google Scholar]
  46. Sears A. E., McGwire B. S., Roizman B. Infection of polarized MDCK cells with herpes simplex virus 1: two asymmetrically distributed cell receptors interact with different viral proteins. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5087–5091. doi: 10.1073/pnas.88.12.5087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Shieh M. T., WuDunn D., Montgomery R. I., Esko J. D., Spear P. G. Cell surface receptors for herpes simplex virus are heparan sulfate proteoglycans. J Cell Biol. 1992 Mar;116(5):1273–1281. doi: 10.1083/jcb.116.5.1273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Sunstrum J. C., Chrisp C. E., Levine M., Glorioso J. C. Pathogenicity of glycoprotein C negative mutants of herpes simplex virus type 1 for the mouse central nervous system. Virus Res. 1988 Aug;11(1):17–32. doi: 10.1016/0168-1702(88)90064-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Superti F., Hauttecoeur B., Morelec M. J., Goldoni P., Bizzini B., Tsiang H. Involvement of gangliosides in rabies virus infection. J Gen Virol. 1986 Jan;67(Pt 1):47–56. doi: 10.1099/0022-1317-67-1-47. [DOI] [PubMed] [Google Scholar]
  50. Söderberg C., Giugni T. D., Zaia J. A., Larsson S., Wahlberg J. M., Möller E. CD13 (human aminopeptidase N) mediates human cytomegalovirus infection. J Virol. 1993 Nov;67(11):6576–6585. doi: 10.1128/jvi.67.11.6576-6585.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Taylor H. P., Cooper N. R. The human cytomegalovirus receptor on fibroblasts is a 30-kilodalton membrane protein. J Virol. 1990 Jun;64(6):2484–2490. doi: 10.1128/jvi.64.6.2484-2490.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Trybala E., Bergström T., Svennerholm B., Jeansson S., Glorioso J. C., Olofsson S. Localization of a functional site on herpes simplex virus type 1 glycoprotein C involved in binding to cell surface heparan sulphate. J Gen Virol. 1994 Apr;75(Pt 4):743–752. doi: 10.1099/0022-1317-75-4-743. [DOI] [PubMed] [Google Scholar]
  53. Trybala E., Svennerholm B., Bergström T., Olofsson S., Jeansson S., Goodman J. L. Herpes simplex virus type 1-induced hemagglutination: glycoprotein C mediates virus binding to erythrocyte surface heparan sulfate. J Virol. 1993 Mar;67(3):1278–1285. doi: 10.1128/jvi.67.3.1278-1285.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Tucker S. P., Thornton C. L., Wimmer E., Compans R. W. Bidirectional entry of poliovirus into polarized epithelial cells. J Virol. 1993 Jan;67(1):29–38. doi: 10.1128/jvi.67.1.29-38.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Vanderplasschen A., Bublot M., Dubuisson J., Pastoret P. P., Thiry E. Attachment of the gammaherpesvirus bovine herpesvirus 4 is mediated by the interaction of gp8 glycoprotein with heparinlike moieties on the cell surface. Virology. 1993 Sep;196(1):232–240. doi: 10.1006/viro.1993.1471. [DOI] [PubMed] [Google Scholar]
  56. Whealy M. E., Robbins A. K., Enquist L. W. Pseudorabies virus glycoprotein gIII is required for efficient virus growth in tissue culture. J Virol. 1988 Jul;62(7):2512–2515. doi: 10.1128/jvi.62.7.2512-2515.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Whealy M. E., Robbins A. K., Tufaro F., Enquist L. W. A cellular function is required for pseudorabies virus envelope glycoprotein processing and virus egress. J Virol. 1992 Jun;66(6):3803–3810. doi: 10.1128/jvi.66.6.3803-3810.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. WuDunn D., Spear P. G. Initial interaction of herpes simplex virus with cells is binding to heparan sulfate. J Virol. 1989 Jan;63(1):52–58. doi: 10.1128/jvi.63.1.52-58.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Zsak L., Sugg N., Ben-Porat T., Robbins A. K., Whealy M. E., Enquist L. W. The gIII glycoprotein of pseudorabies virus is involved in two distinct steps of virus attachment. J Virol. 1991 Aug;65(8):4317–4324. doi: 10.1128/jvi.65.8.4317-4324.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Zsak L., Sugg N., Ben-Porat T. The different interactions of a gIII mutant of pseudorabies virus with several different cell types. J Gen Virol. 1992 Apr;73(Pt 4):821–827. doi: 10.1099/0022-1317-73-4-821. [DOI] [PubMed] [Google Scholar]
  61. Zsak L., Zuckermann F., Sugg N., Ben-Porat T. Glycoprotein gI of pseudorabies virus promotes cell fusion and virus spread via direct cell-to-cell transmission. J Virol. 1992 Apr;66(4):2316–2325. doi: 10.1128/jvi.66.4.2316-2325.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Zuckermann F. A., Mettenleiter T. C., Schreurs C., Sugg N., Ben-Porat T. Complex between glycoproteins gI and gp63 of pseudorabies virus: its effect on virus replication. J Virol. 1988 Dec;62(12):4622–4626. doi: 10.1128/jvi.62.12.4622-4626.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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