Synthesis in Escherichia coli of the reovirus nonstructural protein sigma NS

Abstract

The coding region of reovirus type 3 genomic segment S3, encoding the nonstructural protein sigma NS, was placed under the control of the bacteriophage lambda pL promoter in the Escherichia coli expression plasmid pRC23 (J.C. Lacal, E. Santos, V. Notario, M. Barbacid, S. Yamazaki, H.-F. Kung, C. Seamans, S. McAndrew, and R. Crowl, Proc. Natl. Acad. Sci. USA 81:5305-5309). Derepression of the pL promoter led to the synthesis of a protein of the same molecular weight as sigma NS produced in reovirus-infected L cells. The expressed protein was indistinguishable from authentic sigma NS by peptide mapping with Staphylococcus aureus V8 protease and by immunoblot analysis. Most importantly, the purified protein had nucleic acid-binding properties similar to that previously shown for sigma NS obtained from infected cells. Binding of single-stranded RNAs by recombinant sigma NS protein was inhibited by GTP.