RNA polymerase binding sites isolated from T4DNA: analysis of oligopyrimidine sequences constituting preinitiation and initiation complexes.

Abstract

Oligopyrimidines which contain 5'-hydroxymethylcytosine instead of cytosine were separated by thin layer chromatography. Using this method, the oligopyrimidine pattern of RNA polymerase binding sites, isolated from T4DNA, was evaluated quantitatively. The analysis shows that 1. the RNA polymerase binding sites on T4 DNA obtained under low salt conditions in absence of triphosphates, are A-T-rich as compared with total T4 DNA. 2. The A-T base pairs stand mainly in alternating position. On the average these sequences comprise more than half of the chain length of each binding site, which contains about 8 G-5'-HMC pairs. 3. The sites of binding and the sites of initiation do not show an identical base composition. 4. A mixture of at least 8 different binding istes is isolated under the conditions employed. This figure is in agreement with the number of distinct transcripts synthesized in nitro by E. coli RNA polymerase from T4 DNA. The overall length of these transcripts corresponds to approximately 9% of the T4 genome.