Determination of the DNA conformation of the simian virus 40 (SV40) enhancer in SV40 minichromosomes.
The simian virus 40 (SV40) enhancer contains three 8-bp purine-pyrimidine alternating sequences which are known to adopt the left-handed Z-DNA conformation in vitro. In this paper, we have undertaken the determination of the DNA conformation adopted by these Z-motifs in the SV40 minichromosome. We have analyzed the presence of Z-DNA through the change in linkage which should accompany formation of this left-handed conformation. Our results indicate that, regardless of the precise moment of the viral lytic cycle at which minichromosomes are harvested and the condition of the transfected DNA, either relaxed or negatively supercoiled, none of the three Z motifs of the SV40 enhancer exist to a significant extent as Z-DNA in SV40 minichromosomes. The SV40 enhancer adopts predominantly a right-handed B-DNA conformation in vivo.
Martínez-Balbás
A
A
Grupo de Química Macromolecular, Escuela Técnica Superior de Ingenieros Industriales, Barcelona, Spain.
Casasnovas
J M
JM
Azorín
F
F
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002874:Chromosome Mapping; D004247:DNA / Q000032:analysis / Q000235:genetics; D004278:DNA, Superhelical / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics*; D015246:Deoxyribonuclease EcoRI; D004586:Electrophoresis / Q000379:methods; D004742:Enhancer Elements, Genetic; D005814:Genes, Viral; D008040:Genetic Linkage; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D013539:Simian virus 40 / Q000235:genetics*; D014158:Transcription, Genetic; D015678:Viral Structural Proteins / Q000235:genetics
D004278:DNA, Superhelical; D004279:DNA, Viral; D015678:Viral Structural Proteins; D004247:DNA; D015246:Deoxyribonuclease EcoRI