Sequence-Specific Interaction of R17 Coat Protein with Its Ribonucleic Acid Binding Site

Jannette Carey, Olke C. Uhlenbeck, Vicki Cameron, Pieter L. de Haseth

Research output: Contribution to journalArticle

197 Scopus citations

Abstract

The interaction between phage R17 coat protein and its RNA binding site for translational repression was studied as an example of a sequence-specific RNA-protein interaction. Nuclease protection and selection experiments define the binding site to about 20 contiguous nucleotides which form a hairpin. A nitrocellulose filter retention assay is used to show that the binding between the coat protein and a synthetic 21-nucleotide RNA fragment conforms to a simple bimolecular reaction. Unit stoichiometry and a Kd of about 1 nM are obtained at 2 °C in buffer containing 0.19 M salt. The interaction is highly sequence specific since a variety of RNAs failed to compete with the 21-nucleotide fragment for coat protein binding.

Original languageEnglish (US)
Pages (from-to)2601-2610
Number of pages10
JournalBiochemistry
Volume22
Issue number11
DOIs
StatePublished - Jan 1 1983
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry

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