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PubMed · 8139557

Cellular protein interactions with herpes simplex virus type 1 oriS.

C E Dabrowski·P J Carmillo·P A Schaffer
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Abstract

The herpes simplex virus type 1 (HSV-1) origin of DNA replication, oriS, contains an AT-rich region and three highly homologous sequences, sites I, II, and III, identified as binding sites for the HSV-1 origin-binding protein (OBP). In the present study, interactions between specific oriS DNA sequences and proteins in uninfected cell extracts were characterized. The formation of one predominant protein-DNA complex, M, was demonstrated in gel shift assays following incubation of uninfected cell extracts with site I DNA. The cellular protein(s) that comprises complex M has been designated origin factor I (OF-I). The OF-I binding site was shown to partially overlap the OBP binding site within site I. Complexes with mobilities indistinguishable from that of complex M also formed with site II and III DNAs in gel shift assays. oriS-containing plasmid DNA mutated in the OF-I binding site exhibited reduced replication efficiency in transient assays, demonstrating a role for this site in oriS function. The OF-I binding site is highly homologous to binding sites for the cellular CCAAT DNA-binding proteins. The binding site for the CCAAT protein CP2 was found to compete for OF-I binding to site I DNA. These studies support a model involving the participation of cellular proteins in the initiation of HSV-1 DNA synthesis at oriS.

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AnimalsBase SequenceBinding SitesBinding, CompetitiveConsensus SequenceDNADNA ReplicationDNA-Binding ProteinsHerpesvirus 1, HumanKineticsMolecular Sequence DataMutagenesisOligonucleotide ProbesSequence Homology, Nucleic AcidVero CellsViral ProteinsJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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C E Dabrowski, P J Carmillo, P A Schaffer. 1994. Cellular protein interactions with herpes simplex virus type 1 oriS.. https://doi.org/10.1128/mcb.14.4.2545-2555.1994

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