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Molecular and Cellular Biology, November 1998, p. 6399-6407, Vol. 18, No. 11
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Middle Subunit of Replication Protein A Contacts Growing RNA-DNA Primers in Replicating Simian Virus 40 Chromosomes

Gilad Mass, Tamar Nethanel, and Gabriel Kaufmann*

Department of Biochemistry, Tel Aviv University, Ramat Aviv 69978, Israel

Received 27 March 1998/Returned for modification 26 May 1998/Accepted 6 August 1998

The eukaryotic single-stranded DNA binding protein replication protein A (RPA) participates in major DNA transactions. RPA also interacts through its middle subunit (Rpa2) with regulators of the cell division cycle and of the response to DNA damage. A specific contact between Rpa2 and nascent simian virus 40 DNA was revealed by in situ UV cross-linking. The dynamic attributes of the cross-linked DNA, namely, its size distribution, RNA primer content, and replication fork polarity, were determined. These data suggest that Rpa2 contacts the early DNA chain intermediates synthesized by DNA polymerase alpha -primase (RNA-DNA primers) but not more advanced products. Possible signaling functions of Rpa2 are discussed, and current models of eukaryotic lagging-strand DNA synthesis are evaluated in view of our results.

* Corresponding author. Mailing address: Department of Biochemistry, Tel Aviv University, Ramat Aviv 69978, Israel. Phone: 972-3-640-9067. Fax: 972-3-642-6213. E-mail: kaufmann{at}post.tau.ac.il.

Molecular and Cellular Biology, November 1998, p. 6399-6407, Vol. 18, No. 11
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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Copyright © 1998 by the American Society for Microbiology. All rights reserved.