Bends in SV40 DNA: use of mutagenesis to identify the critical bases involved

Abstract

Five fragments of DNA exhibiting sequence directed bends were isolated from the Simian Virus 40 genome using a two-dimensional polyacrylamide gel fractionation. The bend sites were mapped for each fragment using the circular permutation test. All five sites have multiple, short runs of A residues with helical spacing typical of other bent fragments. Base pairs important for the bends were determined for one fragment by utilizing a random, single base pair mutagenesis. Of 28 mutants with decreased or increased bends, 14 had alterations that could be interpreted to affect the spaced runs of A residues, supporting their role in bends as predicted by the ApA wedge model. One major mutation was not explainable by existing models. The remaining minor mutations may only be due to small, local DNA conformational changes in the surrounding B-DNA.