Summary
We have previously described two serologically related DNA-binding phosphoproteins of different apparent molecular mass (75 kDa and 35 kDa) produced in cottontail rabbit herpesvirus (CTHV)-infected cells. The 75 kDa protein appeared before the 35 kDa protein in the infectious cycle. Here, we extend the characterization of these proteins. Protease V8 fingerprints of methionine-labelled 35 kDa protein showed four major peptide products, three of which comigrated with major peptides from digests of the 75 kDa protein. The fourth peptide, with an estimated mass of 10 kDa, reacted with an antiserum recognizing both proteins. In vitro translation of total or poly A-containing RNA isolated from infected cells at 24 h to 72 h post-infection produced only the 75 kDa protein as measured by immunoprecipitation with anti-75/35 kDa serum, suggesting that the 35 kDa protein is derived from the 75 kDa protein by proteolytic cleavage. Virus-specific RNA obtained by prehybridization to CTHV DNA also produced the 75 kDA protein, confirming its viral origin. The putative gene for the 75 kDa protein was localized to a region on the CTHV DNA restricted byPvuII.
References
Baum EZ, Bebernitz GA, Hulmes JD, Muzithras VP, Jones TR, Gluzman Y (1992) Expression and analysis of the human cytomegalovirus UL80-encoded protease: identification of autoproteolytic site. J Virol 67: 497–506
Bornkamm GW, Delius H, Fleckenstein B, Werner FJ, Mulder C (1976) The structure of herpesvirus saimiri genome: arrangement of heavy and light sequences within the M genome. J Virol 19: 154–161
Bridgen A, Herring AJ, Inglis NS, Reid HW (1989) Preliminary characterization of the alcelaphine herpesvirus 1 genome. J Gen Virol 70: 1141–1150
Bublot M, Lomonte P, Lequarre AS, Albrecht JC, Nicholas J, Fleckenstein B, Pastoret PP, Thyry E (1992) Genetic relationships between bovine herpesvirus 4 and the gammaherpes viruses Epstein-Barr virus and herpesvirus saimiri. Virology 190: 654–665
Cajean-Feroldi C, Laithier M, Foulon T, Sheldrick P (1988) DNA-binding proteins induced by the cottontail rabbit herpesvirus CTHV. J Gen Virol 69: 2277–2289
Cebrian J, Berthelot N, Laithier M (1989) Genome structure of cottontail rabbit herpesvirus. J Virol 63: 523–531
Chirgwin JM, Przbyla AE, Macdonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294–5299.
Cleveland D, Fisher S, Kirshner M, Laemmli U (1977) Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem 252: 1102–1106
Ehlers B, Buhk HJ, Ludwig H (1985) Analysis of bovine cytomegalovirus genome structure: cloning and mapping of the monomeric polyrepetitive DNA unit, and comparison of European and American strains. J Gen Virol 66: 55–68
Efstathiou T, Ho YM, Hall S, Styles CJ, Scott SD, Gompels, UA (1990) Murine herpesvirus 68 is genetically related to the gammaherpesviruses Epstein-Barr virus and herpesvirus saimiri. J Gen Virol 71: 1365–1372
Fleckenstein B, Desrosiers RC (1982) Herpesvirus saimiri and herpesvirus ateles. In: Roizman B (ed) The herpesviruses, Vol 1. Plenum Press, New York, pp 253–332
Foulon T, Cebrian J (1989) Restriction maps for the cottontail rabbit herpesvirus (CTHV) genome. Gene 78: 371–376
Foulon T, Pertuiset B, Cajean-Feroldi C, Sheldrick P (1993) Possible relation between the U54 segment of the CTHV genome and the conserved gene block C rearranged in alpha and gamma herpesvirus genomes. Virus Res 29: 115–124
Fuchs PG, Ruger R, Pfister H, Fleckenstein B (1985) Genome organization of herpesvirus aotus type 2. J Virol 53: 13–18
Liu F, Roizman B (1991a) 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 65: 206–212
Liu F, Roizman B (1991b) The herpes simplex virus 1 gene encoding a protease also contains within its coding domain the gene encoding the more abundant substrate. J Virol 65: 5149–5156
Liu F, Roizman B (1993) Characterization of the protease and other products of aminoterminus-proximal cleavage of the herpes simplex virus 1 UL26 protein. J Virol 67: 1300–1309
Mason PJ, Williams JG (1985) Hybridization in the analysis of recombinant DNA. In: Hames BD, Higgins S (eds) Nucleic acid hybridization. A practical approach. IRL Press, Oxford, pp 128–133
Medveczky P, Kramp WP, Sullivan JL (1984) Circular herpesvirus sylvilagus DNA in spleen cells of experimentally infected cottontail rabbits. J Virol 52: 711–714
Medveczky MM, Geck P, Clarke C, Byrnes J, Sullivan JL, Medveczky PG (1989) Arrangement of repetitive sequences in the genome of herpesvirus sylvilagus. J Virol 63: 1010–1014
Roizman B, Desrosiers RC, Fleckenstein B, Lopez C, Minson AC, Studdert MJ (1992) The family herpesviridae: an update. Arch Virol 123: 425–449
Rouhandeh H, Cohrs R (1987) Cloning and physical mapping of herpesvirus sylvilagus DNA. Gene 61: 131–139
Schwyzer M (1977) Purification of SV 40 T antigen by immune affinity chromatography on staphylococcal protein A-sepharose. INSERM-Colloq 69: 63–68
Stortz J, Ehlers B, Todd WJ, Ludwig H (1984) Bovine cytomegalovirus: identification and differential properties. J Gen Virol 65: 697–706
Welch AR, Wood AS, McNally LM, Cotter RJ, Gibson W (1991) A herpes maturational protease, assemblin: identification of its gene, putative active site domain, and cleavage site. Proc Natl Acad Sci USA 88: 10792–10796
Weinheimer SP, McCann III PJ, O'Boyle II DR, Stevens JT, Boyd BA, Drier DA, Yamanaka GA, DiIanni CL, Deckman IC, Cordingley MG (1993) Autoproteolysis of herpes simplex type 1 protease releases an active catalytic domain found in intermediate capsid particles. J Virol 67: 5813–5822
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Cajean-Feroldi, C., Laithier, M. Partial characterization of two serologically related DNA-binding proteins specified by the cottontail rabbit herpesvirus CTHV. Archives of Virology 140, 1493–1501 (1995). https://doi.org/10.1007/BF01322676
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DOI: https://doi.org/10.1007/BF01322676