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Nucleic Acids Research, 1986, Vol. 14, No. 22 8949-8962
© 1986

Articles

Altered DNA conformations In the gene regulatory region of torsionaliy-stressed SV4O DNA

Lauren lacono-Connors and David Kowalski

Roswell Park Memorial Institute, Department of Molecular and Cellular Biology 666 Elm Street, Buffalo, NY 14263, USA

Received July 24, 1986. Revised October 11, 1986. Accepted October 11, 1986.

We used mung bean nuclease to probe the SV4O genome for DNA unwinding and unpairing. Cleavage occurred at a limited number of specific sites in supercoiled, but not relaxed DNA. The nurber and location of cleavage sites depended upon Mg2+ concentration. Without Mg2+ , cutting occurred mainly in one early denaturation region located 3' to the t antigen gene and within the T antigen gene intron. With Mg2+ , cleavage occurred at a number of alternative sites in the genome. Certain Mg2+ concentrations favored cleavage in the gene regulatory region. These cleavages were mapped at single nucleotide resolution and occurred in both transcriptional enhancers and upstream from the start of major late gene transcription. The cleavages occurred between 5 bp inverted repeat sequences, consistent with the recognition of unusually small cruciform structures.


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