Production of simian virus 40 large tumor antigen in bacteria: altered DNA-binding specificity and dna-replication activity of underphosphorylated large tumor antigen

Mohr, I. J., Gluzman, Y., Fairman, M. P., Strauss, M., McVey, D., Stillman, B., Gerard, R. D. (1989) Production of simian virus 40 large tumor antigen in bacteria: altered DNA-binding specificity and dna-replication activity of underphosphorylated large tumor antigen. Proceedings of the National Academy of Sciences of the United States of America, 86 (17). pp. 6479-83. ISSN 0027-8424

URL: http://www.ncbi.nlm.nih.gov/pubmed/2549538

Abstract

A bacterial expression system was used to produce simian virus 40 large tumor antigen (T antigen) in the absence of the extensive posttranslational modifications that occur in mammalian cells. Wild-type T antigen produced in bacteria retained a specific subset of the biochemical activities displayed by its mammalian counterpart. Escherichia coli T antigen functioned as a helicase and bound to DNA fragments containing either site I or the wild-type origin of replication in a manner identical to mammalian T antigen. However, T antigen purified from E. coli did not efficiently bind to site II, an essential cis element within the simian virus 40 origin of replication. It therefore could not unwind origin-containing plasmids or efficiently replicate simian virus 40 DNA in vitro. The ability of protein phosphorylation to modulate the intrinsic preference of full-length T antigen for either site I or site II is discussed.

Item Type: Paper
Additional Information:
Uncontrolled Keywords: Antigens, Polyomavirus Transforminggenetics Base Sequence DNA Replication DNA Binding Proteinsgenetics Escherichia coli genetics Hela Cellsimmunology Humans Kinetics Molecular Sequence Data Oligonucleotide Probes Plasmids Restriction Mapping Simian virus 40 genetics immunology
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
organism description > bacteria
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date Deposited: 02 Mar 2012 20:01
Last Modified: 20 Jun 2017 20:19
PMCID: PMC297867
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25026

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