Characterization of Genomic Poly(dT-dG) · Poly(dC-dA) Sequences: Structure, Organization, and Conformation

Abstract

Hybridization studies suggest the abundant presence of poly(dT-dG) · poly(dC-dA) (TG-element), a potential Z-DNA sequence, in eucaryotic genomes. We have isolated and characterized TG-elements from different locations in the human genome: from randomly isolated clones, associated with the actin gene family, and linked to another repeated element. The results indicate that the following features are typical of these TG-elements: (i) the elements consist of 20 to 60 base pairs of (dT-dG)_n · (dC-dA)_n, (ii) the sequences characterized in our study were not flanked by direct or inverted repeats, (iii) the sequences are interspersed rather than in satellite blocks, (iv) the elements are not usually associated with other repeated elements, and (v) some of the elements are found near coding sequences or in introns. Studies on the conformation of a genomic TG-element in a supercoiled plasmid indicate several distinct properties of the TG-element: (i) it is in the Z-form only at low ionic strength, (ii) S1 nuclease recognizes its Z-form with a marked preference for one of the B-Z junctions, and (iii) the sensitive region extends for 20 base pairs near the B-Z junction. In contrast to the result with the supercoiled plasmid, S1 nuclease failed to recognize the TG-element in minichromosomes.