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Journal of Virology, October 2003, p. 10651-10657, Vol. 77, No. 19
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.19.10651-10657.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Relationship among Location of T-Antigen-Induced DNA Distortion, Auxiliary Sequences, and DNA Replication Efficiency

Susan Okuley,{dagger} Mindy Call,{ddagger} Tara Mitchell,§ Bugen Hu,|| and Mary E. Woodworth*

Department of Microbiology, Miami University, Oxford, Ohio 45056

Received 2 April 2003/ Accepted 16 June 2003

T-antigen-induced DNA distortion was studied in a series of simian virus 40 (SV40) plasmid constructs whose relative replication efficiency ranges from 0.2 to 36. Bending was detected in the wild-type SV40 regulatory region consisting of three copies of the GC-rich 21-bp repeat but not in constructs with only one or two copies of the 21-bp repeat. In a construct with enhanced replication efficiency, bending occurred in a 69-bp cellular sequence located upstream of a single copy of the 21-bp repeat. Bending occurred both upstream of ori and in the three 21-bp repeats located downstream of ori in a construct with reduced replication efficiency. In a construct with no 21-bp repeats, DNA distortion occurred downstream of ori. The results indicate that SV40 DNA replication is enhanced when the structure of the regulatory region allows the DNA to form a bent structure upstream of the initial movement of the replication fork.

* Corresponding author. Mailing address: Department of Microbiology, Miami University, 32 Pearson Hall, Oxford, OH 45056. Phone: (513) 529-7249. Fax: (513) 529-2431. E-mail: woodwome{at}muohio.edu.

{dagger} Present address: Geneformatics, Inc., San Diego, CA 92121.

{ddagger} Present address: Department of Biology, University of Dayton, Dayton, OH 45469.

§ Present address: Corporate Research and Development, Procter & Gamble, Monroe, Ohio.

|| Present address: Preclinical Development, Human Genome Sciences, Inc., Rockville, MD 20850.

Journal of Virology, October 2003, p. 10651-10657, Vol. 77, No. 19
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.19.10651-10657.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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