Conformational changes in deoxyribonucleic acid during transcription.

Abstract

Circular dichroism (CD) was used to examine changes in secondary structure of calf thymus DNA during in vitro transcription. Formation of a binary complex between DNA and RNA polymerase (nucleoside triphosphate:nucleotidyltransferase, EC 2.7.7.6) did not alter the CD spectrum of the DNA. Alterations in ellipticity in the spectral region between 245 and 300 nm occurred during synthesis of RNA. This change was consistent with a B- to A-like form transition in polynucleotide conformation. The increment of ellipticity consisted of two separate compounds; component I was insensitive to treatment with pancreatic ribonuclease whereas component II was a ribonuclease labile fraction. Cleavage by restriction endonucleases did not produce or significantly alter the ellipticity of transcription. In contrast, between 50% and 60% of the component I ellipticity was sensitive to pancreatic DNase I. The data indicate that component I is a property of DNA and suggest that the alteration in secondary conformation which affects this component extends cooperatively beyond the DNase I insensitive DNA-RNA polymerase complexes.