The linear virion DNA of bovine herpesvirus type 4 (BHV-4) is flanked by tandem repeats designated polyrepetitive DNA (prDNA). To investigate the structure and functional role of the prDNA for cleavage/packaging of progeny viral DNA, the complete nucleotide sequence (2267 bp) of a cloned prDNA unit of BHV-4 was determined. Moreover, the terminal fragments of the genome and the junctions between prDNA and the central unique DNA were analysed. In order to characterize the function of the prDNA of BHV-4, a transient packaging assay was developed. The prDNA has a G+C content of 71.1%. Its structure is composed of numerous internal repeats and every unit contains the conserved sequence of the cleavage/packaging signal. A fragment of 443 bp comprising the cleavage/packaging signal was found to be sufficient for cleavage and encapsidation of replicated concatemeric viral DNA. These results suggest that prDNA is a functionally important region of the genome of BHV-4.
BainesJ. D.,
PoonA. P.,
RovnakJ.,
RoizmanB.1994; The herpes simplex virus 1 UL15 gene encodes two proteins and is required for cleavage of genomic viral DNA. Journal of Virology 68:8118–8124
DeissL. P.,
ChouJ.,
FrenkelN.1986; Functional domains within the a sequence involved in the cleavage-packaging of herpes simplex virus DNA. Journal of Virology 59:605–618
EhlersB.,
BuhkH.-J.,
LudwigH.1985; Analysis of bovine cytomegalovirus genome structure: cloning and mapping of the monomeric polyrepetitive DNA unit, and comparison of European and American strains. Journal of General Virology 66:55–68
MocarskiE. S.,
RoizmanB.1982; Structure and role of the herpes simplex virus DNA termini in inversion, circularization and generation of virion DNA. Cell 31:89–97
RoizmanB.,
SearsA. E.1996; Herpes simplex viruses and their replication. In Virology 3rd edn pp 2231–2296 Edited by
FieldsB. N.,
KnipeD. M.,
HowleyP. M.
Philadelphia: Lippincott–Raven;
SangerF.,
NicklenS.,
CoulsonA. R.1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, USA 74:5463–5467
SkaliterR.,
MakhovA. M.,
GriffithJ. D.,
LehmanI. R.1996; Rolling circle DNA replication by extracts of herpes simplex virus type 1-infected human cells. Journal of Virology 70:1132–1136
SpaeteR. R.,
FrenkelN.1985; The herpes simplex virus amplicon: analyses of cis-acting replication functions. Proceedings of the National Academy of Sciences, USA 82:694–698
ZimmermannJ.,
HammerschmidtW.1995; Structure and role of the terminal repeats of Epstein–Barr virus in processing and packaging of virion DNA. Journal of Virology693147–3155