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Journal of Virology, July 2000, p. 5939-5948, Vol. 74, No. 13
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Reovirus Protein sigma NS Binds in Multiple Copies to Single-Stranded RNA and Shares Properties with Single-Stranded DNA Binding Proteins

Anne Lynn Gillian,1 Stephen C. Schmechel,2 Jonathan Livny,1 Leslie A. Schiff,2 and Max L. Nibert1,*

Department of Biochemistry and Institute for Molecular Virology, The College of Agricultural and Life Sciences, and The Graduate School, University of Wisconsin-Madison, Madison, Wisconsin 53706,1 and Department of Microbiology, University of Minnesota Medical School, Minneapolis, Minnesota 554552

Received 14 December 1999/Accepted 5 April 2000

Reovirus nonstructural protein sigma NS interacts with reovirus plus-strand RNAs in infected cells, but little is known about the nature of those interactions or their roles in viral replication. In this study, a recombinant form of sigma NS was analyzed for in vitro binding to nucleic acids using gel mobility shift assays. Multiple units of sigma NS bound to single-stranded RNA molecules with positive cooperativity and with each unit covering about 25 nucleotides at saturation. The sigma NS protein did not bind preferentially to reovirus RNA over nonreovirus RNA in competition experiments but did bind preferentially to single-stranded over double-stranded nucleic acids and with a slight preference for RNA over DNA. In addition, sigma NS bound to single-stranded RNA to which a 19-base DNA oligonucleotide was hybridized at either end or near the middle. When present in saturative amounts, sigma NS displaced this oligonucleotide from the partial duplex. The strand displacement activity did not require ATP hydrolysis and was inhibited by MgCl2, distinguishing it from a classical ATP-dependent helicase. These properties of sigma NS are similar to those of single-stranded DNA binding proteins that are known to participate in genomic DNA replication, suggesting a related role for sigma NS in replication of the reovirus RNA genome.

* Corresponding author. Mailing address: Institute for Molecular Virology, University of Wisconsin-Madison, 1525 Linden Dr., Madison, WI 53706. Phone: (608) 262-1536. Fax: (608) 262-7414. E-mail: mlnibert{at}facstaff.wisc.edu.

Journal of Virology, July 2000, p. 5939-5948, Vol. 74, No. 13
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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