Formation of triple-stranded DNA at d(GA.TC)n sequences prevents nucleosome assembly and is hindered by nucleosomes.
Simple repeating d(GA.TC)n DNA sequences are frequently found at eukaryotic promoters, and in some cases they have been shown to be nucleosome free in vivo. These sequences show a high degree of structural polymorphism and are capable of adopting several types of non-B-DNA conformations. Here we show that the structural versatility of these sequences affects their ability to be packed into nucleosomes. Nucleosome assembly onto short double-stranded DNA fragments carrying d(GA.TC)n sequences of different length (n = 10 and n = 22) is very efficient. However, when the simple repeating sequence is forming a [CT(GA.TC)] triplex, nucleosome assembly is either prevented, as in the case of the d(GA.TC)22 sequence, or results in the destabilization of the triple-stranded conformation, as in the case of the d(GA.TC)10 sequence. Similarly, formation of triple-stranded DNA is hindered when the sequence is organized as nucleosomes. Efficient formation of triplex DNA occurs only at relatively high ionic strength (0.6 M NaCl), when the nucleosome is partially destabilized, and results in the disruption of the nucleosomal particle. These results indicate that nucleosome assembly and triplex formation are competing processes.
Espinás
M L
ML
Department de Biologia Molecular i Cel.lular, Centre d'Investigació i Desenvolupament, Consejo Superior de Investigaciones Científicas, Jordi Girona Salgado 18-26, 08034 Barcelona, Spain. fambmc@cid.csic.es
Jiménez-García
E
E
Martínez-Balbás
A
A
Azorín
F
F
D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000378:metabolism*; D009994:Osmolar Concentration
D009707:Nucleosomes; C051802:triplex DNA; D004247:DNA; D003850:Deoxyribonuclease I