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In vivo assessment of the Z-DNA-forming potential of d(CA · GT)n and d(CG · GC)n sequences cloned into SV40 minichromosomes

AutorCasasnovas, José María; Ellison, Michael J.; Rodríguez-Campos, Antonio; Martínez-Balbás, Marian ; Azorín, Ferran
Fecha de publicación20-ago-1989
CitaciónJournal of Molecular Biology 208(4):537-549 (1989)
ResumenAlternating repeated d(CA · GT)n and d(CG · GC)n sequences constitute a significant proportion of the simple repeating elements found in eukaryotic genomic DNA. These sequences are known to form left-handed Z-DNA in vitro. In this paper, we have addressed the question of the in vivo determination of the Z-DNA-forming potential of such sequences in eukaryotic chromatin. For this purpose, we have investigated the ability of a d(CA · GT)30 sequence and a d(CG · GC)5 sequence to form left-handed Z-DNA when cloned into simian virus 40 (SV40) minichromosomes at two different positions: the TaqI site, which occurs in the intron of the T-antigen gene, and the HpaII site, which is located in the late promoter region within the SV40 control region. Formation of Z-DNA at the inserted repeated sequences was analyzed through the change in DNA linkage associated with the B to Z transition. Our results indicate that regardless of: (1) the site of insertion (either TaqI or HpaII), (2) the precise moment of the viral lytic cycle (from 12 h to 48 h postinfection) and (3) the condition of incorporation of the SV40 recombinants to the host cells (either as minichromosomes or as naked DNA, relaxed or negatively supercoiled), neither the d(CA · GT)30 nor the d(CG · GC)5 sequence are stable in the left-handed Z-DNA conformation in the SV40 minichromosome. The biological relebance of these results is discussed. © 1989
Versión del editorhttp://dx.doi.org/10.1016/0022-2836(89)90146-0
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