Cis and trans requirements for stable episomal maintenance of the BPV-1 replicator

Piirsoo, M., Ustav, E., Mandel, T., Stenlund, A., Ustav, M. (January 1996) Cis and trans requirements for stable episomal maintenance of the BPV-1 replicator. EMBO J, 15 (1). pp. 1-11. ISSN 0261-4189 (Print)0261-4189 (Linking)

URL: http://www.ncbi.nlm.nih.gov/pubmed/8598191

Abstract

Papillomavirus genomes are maintained as multicopy nuclear plasmids in transformed cells. To address the mechanisms by which the viral DNA is stably propagated in the transformed cells, we have constructed a cell line CH04.15 expressing constitutively the viral proteins E1 and E2, that are required for initiation of viral DNA replication. We show that these viral proteins are necessary and sufficient for stable extrachromosomal replication. Using the cell line CH04.15, we have shown that the bovine papillomavirus-1 (BPV-1) minimal origin of replication (MO) is absolutely necessary, but is not sufficient for stable extrachromosomal replication of viral plasmids. By deletion and insertion analysis, we identified an additional element (minichromosome maintenance element, MME) in the upstream regulatory region of BPV-1 which assures stable replication of the MO-containing plasmids. This element is composed of multiple binding sites for the transcription activator E2. MME appears to function in the absence of replication but requires E1 and E2 proteins for activity. In contrast to, for example, Epstein-Barr virus oriP, stably maintained BPV-1 plasmids are not subject to once-per-cell cycle replication as determined by density labelling experiments. These results indicate that papillomavirus episomal replicators replicate independently of the chromosomal DNA of their hosts.

Item Type: Paper
Uncontrolled Keywords: Animals Base Sequence Bovine papillomavirus 1/ genetics CHO Cells Cricetinae DNA Primers/chemistry DNA Replication DNA, Viral/ biosynthesis DNA-Binding Proteins/genetics/metabolism Gene Expression Regulation, Viral Molecular Sequence Data Plasmids Transcriptional Activation Transfection Viral Proteins/genetics/metabolism Virus Replication
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics
organism description > virus
CSHL Authors:
Communities: CSHL labs > Stenlund lab
Depositing User: Matt Covey
Date: 2 January 1996
Date Deposited: 12 Mar 2013 18:50
Last Modified: 12 Mar 2013 18:50
PMCID: PMC449912
Related URLs:
URI: http://repository.cshl.edu/id/eprint/27771

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