Characteristics of Z-DNA helices formed by imperfect (purine-pyrimidine) sequences in plasmids.
The capacities of three synthetic sequences to adopt left-handed helices were evaluated in recombinant plasmids. The sequences consisted of very short runs of (CG)n (n = 2-4) interspersed with runs of alternating A.T base pairs and/or with regions of non-alternating base pairs. The plasmids were studied by two-dimensional gel electrophoresis to determine the natures of the conformational transitions and their free energies of formation. These results coupled with analyses with chemical (diethyl pyrocarbonate, osmium tetroxide, and bromoacetaldehyde) and enzymatic (S1 nuclease, T7 gene 3 product, and MHhaI) probes indicated that the entire sequence was adopting a left-handed helix in each case. In one of these sequences, Z-DNA formation necessitated the retention of the anti conformation of one of the guanines in a region of non-alternation. In a sequence which contains out-of-phase regions of alternation, our results indicate the formation of a separate left-handed helix in the central (CG)2 region, thus forming two Z-Z junctions. In summary, we conclude that only very short regions of alternating CG are necessary to effect the B to Z transition and that this conformational change can be transmitted through non-alternating regions. A set of empirical rules governing the characteristics of the B to Z transition and the types of left-handed helices in supercoiled plasmids was derived from studies on a systematic series of 17 plasmids.
McLean
M J
MJ
Department of Biochemistry, School of Medicine, University of Alabama, Birmingham 35294.
Lee
J W
JW
Wells
R D
RD
D000079:Acetaldehyde / Q000031:analogs & derivatives; D001483:Base Sequence; D004269:DNA, Bacterial; D004047:Diethyl Pyrocarbonate; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides; D009993:Osmium Tetroxide; D010957:Plasmids
D004269:DNA, Bacterial; D009838:Oligodeoxyribonucleotides; C038235:bromoacetaldehyde; D000079:Acetaldehyde; D004047:Diethyl Pyrocarbonate; D009993:Osmium Tetroxide