RNA Cleavage and Chain Elongation by Escherichia coli DNA-Dependent RNA Polymerase in a Binary Enzyme·RNA Complex

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 91; no. 9; pp. 3784 - 3788
• Main Authors: Altmann, Curtis R., Solow-Cordero, David E., Chamberlin, Michael J.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 26.04.1994; National Academy of Sciences
• Subjects: Active sites; Analytical, structural and metabolic biochemistry; Bacteria; Binding sites; Biochemistry; Biological and medical sciences; Buffer storage; Deoxyribonucleic acid; DNA; Enzymes; Enzymes and enzyme inhibitors; Escherichia coli; Fundamental and applied biological sciences. Psychology; Hybridity; Nucleic acids; Nucleotides; Ribonucleic acid; RNA; Transferases; Nucleotidyltransferases; Enzymatic activity; RNA; Enzyme; Transferases; Escherichia coli; Bacteria; Cleavage; Molecular interaction; Molecular complex; DNA-directed RNA polymerase; Enterobacteriaceae
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.91.9.3784

In the absence of DNA, Escherichia coli RNA polymerase (EC 2.7.7.6) can bind RNA to form an equimolar binary complex with the concomitant release of the σ factor. We show now that E. coli RNA polymerase binds at a region near the 3' terminus of the RNA and that an RNA in such RNA·RNA polymerase complexes undergoes reactions previously thought to be unique to nascent RNA in ternary complexes with DNA. These include GreA/GreB-dependent cleavage of the RNA and elongation by 3'-terminal addition of NMP from NTP. Both of these reactions are inhibited by rifampicin. Hence, by several criteria, the RNA in binary complexes is bound to the polymerase in a manner quite similar to that in ternary complexes. These findings can be explained by a model for the RNA polymerase ternary complex in which the RNA is bound at the 3' terminus through two protein binding sites located up to 10 nt apart. In this model, the stability of RNA binding to the polymerase in the ternary complex is due primarily to its interaction with the protein.