1887

Abstract

Several recent reports implicate sequences at or near the C terminus of the catalytic subunit (POL) of herpes simplex virus type 1 (HSV-1) DNA polymerase in its interaction with the accessory protein UL42. We have investigated further the involvement of this region by three different approaches: anti-idiotype antibodies, a competition ELISA and inhibition of the interaction by peptides. Antibodies raised in rabbits to peptides corresponding to regions of POL all reacted in Western blots with POL. Surprisingly, the sera raised against C-terminal peptides (amino acids 1221 to 1235 and 1224 to 1235) also reacted with UL42. The UL42 reactivity was shown to be due to the presence of anti-idiotype antibodies, providing direct evidence for complementarity of the structure of the extreme C terminus of POL to a region of UL42. To measure the contribution of the C terminus of POL to UL42 binding we developed a competition ELISA using POL, a truncated polymerase lacking the carboxyl-terminal 27 amino acids (POLdl) and UL42. UL42 binding to immobilized POL was inhibited approximately four times more effectively by competition, in solution, with POL than with POLdl, indicating that the C-terminal 27 amino acids of POL are responsible for at least 75 % of the binding energy. A peptide corresponding to these 27 amino acids (residues 1209 to 1235) inhibited both the POL-UL42 interaction and the stimulation of POL by UL42 and did so more effectively than peptides corresponding to amino acids just away from the C terminus (residues 1195 to 1223 and 1177 to 1195).

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1994-11-01
2024-03-28
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References

  1. Atherton E., Sheppard R. C.editors 1989 Solid Phase Peptide Synthesis: A Practical Approach Oxford: IRL Press;
    [Google Scholar]
  2. Bassiri R. M., Dvorak J., Utiger R. D. 1979; Thyrotropic releasing hormone. In Methods of Hormone Radioimmunoassay pp. 46–47 Jaffe B. M., Boehrman H. R. Edited by New York: Academic Press;
    [Google Scholar]
  3. Bio-Mega Inc 1990; Antiherpes tetrapeptide derivatives having a substituted aspartic acid side chain.European patent application number EP/0/41/411/333/AI.
  4. Brown M., Faulkner P. 1977; A plaque assay for nuclear polyhedrosis viruses using a solid overlay. Journal of General Virology 36:361–364
    [Google Scholar]
  5. Challberg M. D. 1991; Herpes simplex virus DNA replication. Seminars in Virology 2:247–256
    [Google Scholar]
  6. Chartrand P., Crumpacker C. S., Schaffer P. A., Wilkie N. M. 1980; Physical and genetic analysis of herpes simplex virus DNA polymerase locus. Virology 103:311–326
    [Google Scholar]
  7. Cleveland W. L., Erlanger B. F. 1986; The auto-anti-idiotype strategy for preparing monoclonal antibodies to receptor combining sites. Methods in Enzymology 121:92–107
    [Google Scholar]
  8. Clement I., Sjoberg B. M., Huang Y. C. 1991; Carboxyterminal peptides as probes for Escherichia coli ribonucleotide reductase subunit interaction: kinetic analysis of inhibition studies. Biochemistry 30:5164–5171
    [Google Scholar]
  9. Climent I., Sjoberg B. M., Huang Y. C. 1992; Site-directed mutagenesis and deletion of the carboxyl terminus of Escherichia coli ribonucleotide reductase protein R2. Effects on catalytic activity and subunit interaction. Biochemistry 31:4801–1807
    [Google Scholar]
  10. Coen D. M., Aschman D. P., Gelep P. T., Retondo M. J., Weller S. K., Schaffer P. A. 1984; Fine mapping and molecular cloning of mutations in the herpes simplex virus DNA polymerase locus. Journal of Virology 49:236–247
    [Google Scholar]
  11. Cohen E. A., Gaudreau P., Brazeau P., Langelier Y. 1986; Specific inhibition of herpesvirus ribonucleotide reductase by a nonapeptide derived from the carboxy-terminus of subunit 2. Nature; London: 321441–443
    [Google Scholar]
  12. Conner J., MacFarlane J., Lankinen H., Marsden H. 1992; The unique N terminus of the herpes simplex virus type 1 large subunit is not required for ribonucleotide reductase activity. Journal of General Virology 73:103–112
    [Google Scholar]
  13. Conner J., Furlong J., Murray J., Meighan M., Cross A., Marsden H., Clements J. B. 1993a; Herpes simplex virus type 1 large subunit: regions of the protein essential for subunit interaction and dimerisation. Biochemistry 32:13673–13680
    [Google Scholar]
  14. Conner J., Marsden H., Clements J. B. 1993b; Ribonucleotide reductase of herpesviruses. Reviews in Medical Virology 3:1–10
    [Google Scholar]
  15. Crute J. J., Tsurumi T., Zhu L., Weller S. K., Olivo P. D., Challberg M. D., Mocarski E. S., Lehman I. R. 1989; Herpes simplex virus 1 helicase-primase: a complex of three herpes-encoded gene products. Proceedings of the National Academy of Sciences, U.S.A 86:2186–2189
    [Google Scholar]
  16. Digard P., Coen D. M. 1990; A novel functional domain of an a like DNA polymerase. The binding site on the herpes simplex virus polymerase for the viral UL42 protein. Journal of Biological Chemistry 265:17393–17396
    [Google Scholar]
  17. Digard P., Bebrin W. R., Weisshart K., Coen D. M. 1993a; The extreme C terminus of herpes simplex virus DNA polymerase is crucial for functional interaction with processivity factor UL42 and for viral replication. Journal of Virology 67:398–406
    [Google Scholar]
  18. Digard P., Chow C. S., Pirrit L., Coen D. M. 1993b; Functional analysis of the herpes simplex virus UL42 protein. Journal of Virology 67:1159–1168
    [Google Scholar]
  19. Dorsky D. I., Crumpacker C. S. 1988; Expression of herpes simplex virus type 1 DNA polymerase gene by in vitro translation and effects of gene deletions on activity. Journal of Virology 62:3224–3232
    [Google Scholar]
  20. Dutia B. M., Frame M. C., Subak-Sharpe J. H., Clark W. N., Marsden H. S. 1986; Specific inhibition of herpesvirus ribonucleotide reductase by synthetic peptides. Nature; London: 321439–441
    [Google Scholar]
  21. Gallo M. L., Jackwood D. H., Murphy M., Marsden H. S., Parris D. S. 1988; Purification of the herpes simplex virus type 1 65-kilodalton DNA-binding protein: properties of the protein and evidence of its association with the virus-encoded DNA polymerase. Journal of Virology 62:2874–2883
    [Google Scholar]
  22. Gallo M. L., Dorsky D. I., Crumpacker C. S., Parris D. S. 1989; The essential 65-kilodalton DNA-binding protein of herpes simplex virus stimulates the virus-encoded DNA polymerase. Journal of Virology 63:5023–5029
    [Google Scholar]
  23. Gao M., DiTusa S. F., Cordingley M. G. 1993; The C-terminal third of UL42, a HSV-1 DNA replication protein, is dispensable for viral growth. Virology 194:647–653
    [Google Scholar]
  24. Gottlieb J., Marcy A. I., Coen D. M., Challberg M. D. 1990; The herpes simplex virus type 1 UL42 gene product: a subunit of DNA polymerase that functions to increase processivity. Journal of Virology 64:5976–5987
    [Google Scholar]
  25. Hernandez T. R., Lehman I. R. 1990; Functional interaction between the herpes simplex-1 DNA polymerase and UL42 protein. Journal of Biological Chemistry 265:11227–11232
    [Google Scholar]
  26. Izadyar L., Friboulet A., Remy M. L., Roseto A., Thomas D. 1993; Monoclonal anti-idiotypic antibodies as functional internal images of enzyme active sites: production of a catalytic antibody with a cholinesterase activity. Proceedings of the National Academy of Sciences, U.S.A 90:8876–8880
    [Google Scholar]
  27. Kitts P. A., Ayres M. D., Possee R. D. 1990; Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors. Nucleic Acids Research 18:5667–5672
    [Google Scholar]
  28. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227:680–685
    [Google Scholar]
  29. McLean G. W., Owsianka A. M., Subak-Sharpe J. K., Marsden H. S. 1991; Generation of anti-peptide and anti-protein sera: effect of peptide presentation on immunogenicity. Journal of Immuno logical Methods 137:149–157
    [Google Scholar]
  30. Marcy A. I., Olivo P., Challberg M. D., Coen D. M. 1990; Enzymatic activities of overexpressed herpes simplex virus DNA polymerase from recombinant baculovirus-infected insect cells. Nucleic Acids Research 18:1207–1215
    [Google Scholar]
  31. Marsden H. S. 1992; Disruption of protein-subunit interactions. Seminars in Virology 3:67–75
    [Google Scholar]
  32. Matsuura Y., Possee R. D., Overton H. A., Bishop D. H. L. 1987; Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. Journal of General Virology 68:1233–1250
    [Google Scholar]
  33. Monahan S. J., Barlam T. F., Crumpacker C. S., Parris D. S. 1993; Two regions of the herpes simplex virus type 1 UL42 protein are required for its functional interaction with the viral DNA polymerase. Journal of Virology 67:5922–5931
    [Google Scholar]
  34. Owsianka A. M., Hart G., Murphy M., Gottlieb J., Boehme R., Challberg M., Marsden H. S. 1993; Inhibition of herpes simplex virus type 1 DNA polymerase activity by peptides from the UL42 accessory protein is largely nonspecific. Journal of Virology 67:258–264
    [Google Scholar]
  35. Parris D. S., Cross A., Haarr L., Orr A., Frame M. C., Murphy M., McGeoch D. J., Marsden H. S. 1988; Identification of the gene encoding the 65-kilodalton DNA binding protein of herpes simplex virus type 1. Journal of Virology 62:818–825
    [Google Scholar]
  36. Purifoy D. J. M., Powell K. L. 1977; Herpes simplex virus DNA polymerases as the site of phosphonoacetate sensitivity: temperature sensitive mutants. Journal of Virology 24:470–477
    [Google Scholar]
  37. Schenk P., Pietschmann S., Gelderblom H., Pauli G., Ludwig H. 1988; Monoclonal antibodies against herpes simplex virus type 1-infected nuclei defining and localizing the ICP8 protein, 65K DNA-binding protein and polypeptides of the ICP35 family. Journal of General Virology 69:99–111 corrigendum 967
    [Google Scholar]
  38. Schick M. R., Kennedy R. C. 1989; Production and characterisation of anti-idiotypic antibody reagents. Methods in Enzymology 178:36–48
    [Google Scholar]
  39. Stow N. D. 1992; Herpes simplex virus type 1 origin-dependent DNA replication in insect cells using recombinant baculoviruses. Journal of General Virology 73:313–321
    [Google Scholar]
  40. Stow N. D. 1993; Sequences at the C-terminus of the herpes simplex virus type 1 UL30 protein are dispensible for DNA polymerase activity but not for viral origin-dependent DNA replication. Nucleic Acids Research 21:87–92
    [Google Scholar]
  41. Tam J. P. 1988; Synthetic peptide vaccine design: synthesis and properties of a high-density multiple antigenic peptide system. Proceedings of the National Academy of Sciences, U.S.A 85:5409–5413
    [Google Scholar]
  42. Tenney D. J., Hurlburt W. W., Bifano M., Stevens J. T., Micheletti P. A., Hamatake R. K., Cordingley M. G. 1993a; Deletions of the carboxy terminus of herpes simplex virus type 1 UL42 define a conserved amino-terminal functional domain. Journal of Virology 67:1959–1966
    [Google Scholar]
  43. Tenney D. J., Micheletti P. A., Stevens J. T., Hamatake R. K., Matthews J. T., Snachez A. R., Hurlburt W. W., Bifano M., Cordingley M. G. 1993b; Mutations in the C terminus of herpes simplex virus type 1 DNA polymerase can affect binding and stimulation by its accessory protein UL42 without affecting basal polymerase activity. Journal of Virology 67:543–547
    [Google Scholar]
  44. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, U.S.A 76:4350–4354
    [Google Scholar]
  45. Vaughan P. J., Purifoy D. J. M., Powell K. L. 1985; DNA-binding protein associated with herpes simplex virus DNA polymerase. Journal of Virology 53:501–508
    [Google Scholar]
  46. Weller S. K. 1991; Genetic analysis of HSV genes required for genome replication. In Herpesvirus Transcription and Its Regulation pp. 105–135 Wagner E. K. Edited by Boca Raton: CRC Press;
    [Google Scholar]
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