trans-acting requirements for replication of Epstein-Barr virus ori-Lyt
| Autor: | S. D. Hayward, E. D. Fixman, Gary S. Hayward |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
DNA Replication
Transcriptional Activation Herpesvirus 4 Human viruses Molecular Sequence Data Restriction Mapping Immunology Eukaryotic DNA replication DNA Primase Genome Viral Transfection Origin of replication Pre-replication complex Polymerase Chain Reaction Microbiology DNA replication factor CDT1 Open Reading Frames Replication factor C Control of chromosome duplication Sequence Homology Nucleic Acid Virology Animals Simplexvirus Amino Acid Sequence Antigens Viral Vero Cells Base Sequence biology DNA replication RNA Nucleotidyltransferases biochemical phenomena metabolism and nutrition Cosmids Molecular biology Epstein-Barr Virus Nuclear Antigens Oligodeoxyribonucleotides Insect Science DNA Viral biology.protein Origin recognition complex Research Article Plasmids |
| Zdroj: | Journal of Virology. 66:5030-5039 |
| ISSN: | 1098-5514 0022-538X |
| DOI: | 10.1128/jvi.66.8.5030-5039.1992 |
| Popis: | Epstein-Barr virus (EBV) utilizes a completely different mode of DNA replication during the lytic cycle than that employed during latency. The latency origin of replication, ori-P, which functions in the replication of the latent episomal form of the EBV genome, requires only a single virally encoded protein, EBNA-1, for its activity. During the lytic cycle, a separate origin, ori-Lyt, is utilized. Relatively little is known about the trans-acting proteins involved in ori-Lyt replication. We established a cotransfection-replication assay to identify EBV genes whose products are required for replication of ori-Lyt. In this assay, a BamHI-H plasmid containing ori-Lyt was replicated in Vero cells cotransfected with the BamHI-H target, the three EBV lytic-cycle transactivators Zta, Rta, and Mta, and the EBV genome provided in the form of a set of six overlapping cosmid clones. By removing individual cosmids from the cotransfection mixture, we found that only three of the six cosmids were necessary for ori-Lyt replication. Subcloning of the essential cosmids led to the identification of six EBV genes that encode replication proteins. These genes and their functions (either known or predicted on the basis of sequence comparison with herpes simplex virus) are BALF5, the DNA polymerase; BALF2, the single-stranded DNA-binding protein homolog; BMRF1, the DNA polymerase processivity factor; BSLF1 and BBLF4, the primase and helicase homologs; and BBLF2/3, a potential homolog of the third component of the helicase-primase complex. In addition, ori-Lyt replication in this cotransfection assay was also dependent on one or more genes provided by the EBV SalI-F fragment and on the three lytic-cycle transactivators Zta, Rta, and Mta. |
| Databáze: | OpenAIRE |
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