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Nucleic Acids Research, 1989, Vol. 17, No. 16 6515-6522
© 1989

MOLECULAR BIOLOGY

Tet repressor binding induced curvature of tet operator DNA

Kariheinz Tovar and Wolfgang Hillen*

Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie und Biochemie der Friedrich-Alexander Universitat Erlangen-Nüraberg Staudtstrasse 5, 8520 Erlangen, FRG

*To whom correspondence should be addressed

Received June 8, 1989. Accepted July 25, 1989.

Tet repressor dimer binds to two tet operator sites spaced by 30 bp in the Tn10 encoded tet regulatory DNA. The effect of repressor binding on the gel mobility of circular permutated DNA fragments containing either one or both operator sequences is reported. The EcoRI induced bending of DNA is used to compare the results with other protein binding induced structural perturbations of DNA. Tet repressor bends a DNA fragment with a single tet operator to an angle of 42° ± 7°. The apparent bend angle of DNA fragments containing the tandem tet operator arrangement occupied by two Tet repressor dimers turns out to be 52° ± 9°. These results are interpreted with respect to the end to end distances of the bent DNA fragments. They indicate that either the intervening tet regulatory DNA between the operators or the bound operator sequences themselves contain additional perturbations from the canonical B-DNA structure. This finding is discussed in the light of previously obtained results from CD, neutron scattering, and electrooptical studies.


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