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Nucleic Acids Research, 1984, Vol. 12, No. 2 1265-1276
© 1984

MOLECULAR BIOLOGY

DNA unwinding ability of Xenopus transcription factor A

Jay S. Hanas, Daniel F. Bogenhagen and Cheng-Wen Wu

Department of Pharmacological Sciences, School of Medicine, State University of New York Stony Brook, NY 11794, USA

Received September 26, 1983. Revised December 9, 1983. Accepted December 9, 1983.

The duplex DNA unwinding ability of Xenopus transcription factor A was examined by treatment of plasmid-protein complexes with calf thymus topoisomerase I and resolution of the topoisomer distributions by agarose gel electrophoresis. Factor A binding to plasmids does result in a small degree of unwinding as evidenced by a shift in topoisomer distributions upon relaxation. Slightly more unwinding by factor A was observed on a specific plasmid, containing four tandemly repeated 5S RNA genes, when compared to a non-specific plasmid of nearly identical size containing no 5S specific sequences. The amount of unwinding observed on the specific plasmid, about one base-pair per gene, suggests that unwinding of the DNA duplex is not an important aspect of binding of factor A to the 5S RNA gene.


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