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. 1993 Mar;67(3):1357–1364. doi: 10.1128/jvi.67.3.1357-1364.1993

Mutations in herpes simplex virus type 1 genes encoding VP5 and VP23 abrogate capsid formation and cleavage of replicated DNA.

P Desai 1, N A DeLuca 1, J C Glorioso 1, S Person 1
PMCID: PMC237505  PMID: 8382300

Abstract

The herpes simplex virus type 1 capsid is composed of seven capsid proteins which are termed VP5, VP19c, VP21, VP22a, VP23, VP24, and VP26. Major capsid protein VP5 is encoded by the gene UL19. UL18, whose transcript is 3' coterminal with that of VP5, specifies capsid protein VP23. Vero cell lines have been isolated that are transformed with either the BglII N (UL19) or EcoRI G (UL16 to UL21) fragment of KOS. These cell lines, selected for the ability to support the replication of a temperature-sensitive VP5 mutant, were used to isolate VP5 and VP23 null mutants. The mutations in VP5 (K5 delta Z) and VP23 (K23Z) were generated by insertion of the lacZ gene at the beginning of the coding sequences of the genes. Both mutants failed to form plaques on the nonpermissive cell line, and therefore, VP23, like VP5, is an essential gene product for virus replication. Both mutants expressed wild-type levels of infected-cell proteins upon infection of permissive and nonpermissive cell lines. However, the VP5 (150-kDa) and VP23 (33-kDa) polypeptides were absent in lysates prepared from K5 delta Z- and K23Z-infected Vero cells, respectively. No capsid structures were observed by electron microscopic analysis of thin sections of K5 delta Z- and K23Z-infected Vero cells. Following sedimentation of lysates from cells infected by the mutants, capsid proteins were not observed in the fractions where capsids normally sediment. The amounts of DNA replicated in the VP5 and VP23 mutant and in KOS-infected Vero cells were the same as in permissive cells. However, genomic ends were not evident in Vero cells infected with the mutants, suggesting that the DNA remains in concatemers and is not processed into unit length genomes.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Addison C., Rixon F. J., Palfreyman J. W., O'Hara M., Preston V. G. Characterisation of a herpes simplex virus type 1 mutant which has a temperature-sensitive defect in penetration of cells and assembly of capsids. Virology. 1984 Oct 30;138(2):246–259. doi: 10.1016/0042-6822(84)90349-0. [DOI] [PubMed] [Google Scholar]
  2. Addison C., Rixon F. J., Preston V. G. Herpes simplex virus type 1 UL28 gene product is important for the formation of mature capsids. J Gen Virol. 1990 Oct;71(Pt 10):2377–2384. doi: 10.1099/0022-1317-71-10-2377. [DOI] [PubMed] [Google Scholar]
  3. Baker T. S., Newcomb W. W., Booy F. P., Brown J. C., Steven A. C. Three-dimensional structures of maturable and abortive capsids of equine herpesvirus 1 from cryoelectron microscopy. J Virol. 1990 Feb;64(2):563–573. doi: 10.1128/jvi.64.2.563-573.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Booy F. P., Newcomb W. W., Trus B. L., Brown J. C., Baker T. S., Steven A. C. Liquid-crystalline, phage-like packing of encapsidated DNA in herpes simplex virus. Cell. 1991 Mar 8;64(5):1007–1015. doi: 10.1016/0092-8674(91)90324-r. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Braun D. K., Batterson W., Roizman B. Identification and genetic mapping of a herpes simplex virus capsid protein that binds DNA. J Virol. 1984 May;50(2):645–648. doi: 10.1128/jvi.50.2.645-648.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cai W. Z., Person S., DebRoy C., Gu B. H. 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. doi: 10.1016/0022-2836(88)90639-0. [DOI] [PubMed] [Google Scholar]
  7. Chakrabarti S., Brechling K., Moss B. Vaccinia virus expression vector: coexpression of beta-galactosidase provides visual screening of recombinant virus plaques. Mol Cell Biol. 1985 Dec;5(12):3403–3409. doi: 10.1128/mcb.5.12.3403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen G. H., Ponce de Leon M., Diggelmann H., Lawrence W. C., Vernon S. K., Eisenberg R. J. Structural analysis of the capsid polypeptides of herpes simplex virus types 1 and 2. J Virol. 1980 May;34(2):521–531. doi: 10.1128/jvi.34.2.521-531.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Costa R. H., Cohen G., Eisenberg R., Long D., Wagner E. Direct demonstration that the abundant 6-kilobase herpes simplex virus type 1 mRNA mapping between 0.23 and 0.27 map units encodes the major capsid protein VP5. J Virol. 1984 Jan;49(1):287–292. doi: 10.1128/jvi.49.1.287-292.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DeLuca N. A., McCarthy A. M., Schaffer P. A. Isolation and characterization of deletion mutants of herpes simplex virus type 1 in the gene encoding immediate-early regulatory protein ICP4. J Virol. 1985 Nov;56(2):558–570. doi: 10.1128/jvi.56.2.558-570.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Furlong D. Direct evidence for 6-fold symmetry of the herpesvirus hexon capsomere. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2764–2766. doi: 10.1073/pnas.75.6.2764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gibson W., Roizman B. Proteins specified by herpes simplex virus. 8. Characterization and composition of multiple capsid forms of subtypes 1 and 2. J Virol. 1972 Nov;10(5):1044–1052. doi: 10.1128/jvi.10.5.1044-1052.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  14. Heilman C. J., Jr, Zweig M., Stephenson J. R., Hampar B. Isolation of a nucleocapsid polypeptide of herpes simplex virus types 1 and 2 possessing immunologically type-specific and cross-reactive determinants. J Virol. 1979 Jan;29(1):34–42. doi: 10.1128/jvi.29.1.34-42.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lemaster S., Roizman B. Herpes simplex virus phosphoproteins. II. Characterization of the virion protein kinase and of the polypeptides phosphorylated in the virion. J Virol. 1980 Sep;35(3):798–811. doi: 10.1128/jvi.35.3.798-811.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Liu F. Y., Roizman B. The herpes simplex virus 1 gene encoding a protease also contains within its coding domain the gene encoding the more abundant substrate. J Virol. 1991 Oct;65(10):5149–5156. doi: 10.1128/jvi.65.10.5149-5156.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liu F. Y., Roizman B. The promoter, transcriptional unit, and coding sequence of herpes simplex virus 1 family 35 proteins are contained within and in frame with the UL26 open reading frame. J Virol. 1991 Jan;65(1):206–212. doi: 10.1128/jvi.65.1.206-212.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McGeoch D. J., Dalrymple M. A., Davison A. J., Dolan A., Frame M. C., McNab D., Perry L. J., Scott J. E., Taylor P. The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol. 1988 Jul;69(Pt 7):1531–1574. doi: 10.1099/0022-1317-69-7-1531. [DOI] [PubMed] [Google Scholar]
  19. Newcomb W. W., Brown J. C., Booy F. P., Steven A. C. Nucleocapsid mass and capsomer protein stoichiometry in equine herpesvirus 1: scanning transmission electron microscopic study. J Virol. 1989 Sep;63(9):3777–3783. doi: 10.1128/jvi.63.9.3777-3783.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Newcomb W. W., Brown J. C. Structure of the herpes simplex virus capsid: effects of extraction with guanidine hydrochloride and partial reconstitution of extracted capsids. J Virol. 1991 Feb;65(2):613–620. doi: 10.1128/jvi.65.2.613-620.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Newcomb W. W., Brown J. C. Use of Ar+ plasma etching to localize structural proteins in the capsid of herpes simplex virus type 1. J Virol. 1989 Nov;63(11):4697–4702. doi: 10.1128/jvi.63.11.4697-4702.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Person S., Knowles R. W., Read G. S., Warner S. C., Bond V. C. Kinetics of cell fusion induced by a syncytia-producing mutant of herpes simplex virus type I. J Virol. 1975 Jan;17(1):183–190. doi: 10.1128/jvi.17.1.183-190.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pertuiset B., Boccara M., Cebrian J., Berthelot N., Chousterman S., Puvion-Dutilleul F., Sisman J., Sheldrick P. Physical mapping and nucleotide sequence of a herpes simplex virus type 1 gene required for capsid assembly. J Virol. 1989 May;63(5):2169–2179. doi: 10.1128/jvi.63.5.2169-2179.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Preston V. G., Coates J. A., Rixon F. J. Identification and characterization of a herpes simplex virus gene product required for encapsidation of virus DNA. J Virol. 1983 Mar;45(3):1056–1064. doi: 10.1128/jvi.45.3.1056-1064.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Preston V. G., Rixon F. J., McDougall I. M., McGregor M., al Kobaisi M. F. Processing of the herpes simplex virus assembly protein ICP35 near its carboxy terminal end requires the product of the whole of the UL26 reading frame. Virology. 1992 Jan;186(1):87–98. doi: 10.1016/0042-6822(92)90063-u. [DOI] [PubMed] [Google Scholar]
  26. Rixon F. J., Cross A. M., Addison C., Preston V. G. The products of herpes simplex virus type 1 gene UL26 which are involved in DNA packaging are strongly associated with empty but not with full capsids. J Gen Virol. 1988 Nov;69(Pt 11):2879–2891. doi: 10.1099/0022-1317-69-11-2879. [DOI] [PubMed] [Google Scholar]
  27. Rixon F. J., Davison M. D., Davison A. J. Identification of the genes encoding two capsid proteins of herpes simplex virus type 1 by direct amino acid sequencing. J Gen Virol. 1990 May;71(Pt 5):1211–1214. doi: 10.1099/0022-1317-71-5-1211. [DOI] [PubMed] [Google Scholar]
  28. Schaffer P. A., Aron G. M., Biswal N., Benyesh-Melnick M. Temperature-sensitive mutants of herpes simplex virus type 1: isolation, complementation and partial characterization. Virology. 1973 Mar;52(1):57–71. doi: 10.1016/0042-6822(73)90398-x. [DOI] [PubMed] [Google Scholar]
  29. Schaffer P. A., Brunschwig J. P., McCombs R. M., Benyesh-Melnick M. Electron microscopic studies of temperature-sensitive mutants of herpes simplex virus type 1. Virology. 1974 Dec;62(2):444–457. doi: 10.1016/0042-6822(74)90406-1. [DOI] [PubMed] [Google Scholar]
  30. Schrag J. D., Prasad B. V., Rixon F. J., Chiu W. Three-dimensional structure of the HSV1 nucleocapsid. Cell. 1989 Feb 24;56(4):651–660. doi: 10.1016/0092-8674(89)90587-4. [DOI] [PubMed] [Google Scholar]
  31. Sherman G., Bachenheimer S. L. Characterization of intranuclear capsids made by ts morphogenic mutants of HSV-1. Virology. 1988 Apr;163(2):471–480. doi: 10.1016/0042-6822(88)90288-7. [DOI] [PubMed] [Google Scholar]
  32. Sherman G., Bachenheimer S. L. DNA processing in temperature-sensitive morphogenic mutants of HSV-1. Virology. 1987 Jun;158(2):427–430. doi: 10.1016/0042-6822(87)90214-5. [DOI] [PubMed] [Google Scholar]
  33. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  34. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  35. Steven A. C., Roberts C. R., Hay J., Bisher M. E., Pun T., Trus B. L. Hexavalent capsomers of herpes simplex virus type 2: symmetry, shape, dimensions, and oligomeric status. J Virol. 1986 Feb;57(2):578–584. doi: 10.1128/jvi.57.2.578-584.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stow N. D., Wilkie N. M. An improved technique for obtaining enhanced infectivity with herpes simplex virus type 1 DNA. J Gen Virol. 1976 Dec;33(3):447–458. doi: 10.1099/0022-1317-33-3-447. [DOI] [PubMed] [Google Scholar]
  37. Vernon S. K., Ponce de Leon M., Cohen G. H., Eisenberg R. J., Rubin B. A. Morphological components of herpesvirus. III. Localization of herpes simplex virus type 1 nucleocapsid polypeptides by immune electron microscopy. J Gen Virol. 1981 May;54(Pt 1):39–46. doi: 10.1099/0022-1317-54-1-39. [DOI] [PubMed] [Google Scholar]
  38. WILDY P., RUSSELL W. C., HORNE R. W. The morphology of herpes virus. Virology. 1960 Oct;12:204–222. doi: 10.1016/0042-6822(60)90195-1. [DOI] [PubMed] [Google Scholar]
  39. Weller S. K., Carmichael E. P., Aschman D. P., Goldstein D. J., Schaffer P. A. Genetic and phenotypic characterization of mutants in four essential genes that map to the left half of HSV-1 UL DNA. Virology. 1987 Nov;161(1):198–210. doi: 10.1016/0042-6822(87)90186-3. [DOI] [PubMed] [Google Scholar]
  40. Zweig M., Heilman C. J., Jr, Hampar B. Identification of disulfide-linked protein complexes in the nucleocapsids of herpes simplex virus type 2. Virology. 1979 Apr 30;94(2):442–450. doi: 10.1016/0042-6822(79)90474-4. [DOI] [PubMed] [Google Scholar]
  41. al-Kobaisi M. F., Rixon F. J., McDougall I., Preston V. G. The herpes simplex virus UL33 gene product is required for the assembly of full capsids. Virology. 1991 Jan;180(1):380–388. doi: 10.1016/0042-6822(91)90043-b. [DOI] [PubMed] [Google Scholar]

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