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Nucleic Acids Research, 1990, Vol. 18, No. 20 6025-6029
© 1990

CHEMISTRY

Thiophosphate interference experiments locate phosphates important for the hammerhead RNA self-cleavage reaction

Duane E. Ruffner and Olke C. Uhlenbeck*

Department of Chemistry and Biochemistry, Campus Box 215, University of Colorado Boulder, CO, 80309, USA

*To whom correspondence should be addressed

Received June 29, 1990. Revised September 23, 1990. Accepted September 23, 1990.

A hammerhead domain of less than 50 nucleotides is responsible for a self-cleavage reaction in the replication of plant RNA pathogens. The hammerhead is composed of three helices joining at a central conserved core of 11 single stranded nucleotides. The core is believed to fold into a tertiary structure that provides functional groups for catalysis and to coordinate one or more divalent metal ions. In this study we use a phosphorothioate substitution interference assay to identify four phosphates in the conserved core which also play a role in the self-cleavage reaction.


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