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Journal of Virology, August 1999, p. 6424-6429, Vol. 73, No. 8
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Use of DNA, RNA, and Chimeric Templates by a Viral RNA-Dependent RNA Polymerase: Evolutionary Implications for the Transition from the RNA to the DNA World

Robert W. Siegel,1,dagger Laurent Bellon,2 Leonid Beigelman,2 and C. Cheng Kao1,*

Department of Biology, Indiana University, Bloomington, Indiana 47405,1 and Department of Chemistry & Biochemistry, Ribozyme Pharmaceuticals Inc., Boulder, Colorado 803092

Received 3 December 1998/Accepted 25 April 1999

All polynucleotide polymerases have a similar structure and mechanism of catalysis, consistent with their evolution from one progenitor polymerase. Viral RNA-dependent RNA polymerases (RdRp) are expected to have properties comparable to those from this progenitor and therefore may offer insight into the commonalities of all classes of polymerases. We examined RNA synthesis by the brome mosaic virus RdRp on DNA, RNA, and hybrid templates and found that precise initiation of RNA synthesis can take place from all of these templates. Furthermore, initiation can take place from either internal or penultimate initiation sites. Using a template competition assay, we found that the BMV RdRp interacts with DNA only three- to fourfold less well than it interacts with RNA. Moreover, a DNA molecule with a ribonucleotide at position -11 relative to the initiation nucleotide was able to interact with RdRp at levels comparable to that observed with RNA. These results suggest that relatively few conditions were needed for an ancestral RdRp to replicate DNA genomes.

* Corresponding author. Mailing address: Department of Biology, Indiana University, Jordan Hall 138, Bloomington, IN 47405. Phone: (812) 855-7959. Fax: (812) 855-6705. E-mail: ckao{at}bio.indiana.edu.

dagger Present address: Life Sciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545.

Journal of Virology, August 1999, p. 6424-6429, Vol. 73, No. 8
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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