TRIPLE-HELIX DNA ALTERS NUCLEOSOMAL HISTONE-DNA INTERACTIONS AND ACTS AS A NUCLEOSOME BARRIER

被引:47
作者
WESTIN, L
BLOMQUIST, P
MILLIGAN, JF
WRANGE, O
机构
[1] KAROLINSKA INST, MED NOBEL INST, DEPT MOLEC & CELLULAR BIOL, GENET MOLEC LAB, S-17177 STOCKHOLM, SWEDEN
[2] GILEAD SCI, FOSTER CITY, CA 94404 USA
基金
美国国家科学基金会;
关键词
D O I
10.1093/nar/23.12.2184
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Oligonucleotides which form triple helical complexes on double-stranded DNA have been previously reported to selectively inhibit transcription both in vitro and in vivo by physically blocking RNA polymerase or transcription factor access to the DNA template. Here we show that a 16mer oligonucleotide, which forms triple helix DNA by binding to a 16 bp homopurine segment, alters the formation of histone-DNA contacts during in vitro nucleosome reconstitution. This effect was DNA sequence-specific and required the oligonucleotide to be present during in vitro nucleosome reconstitution. Binding of the triple helix oligonucleotide on a 199 bp mouse mammary tumour virus promoter DNA fragment with a centrally located tripler DNA resulted in interruption of histone-DNA contacts flanking the tripler DNA segment. When nucleosome reconstitution is carried out on a longer, 279 bp DNA fragment with an asymmetrically located tripler site, nucleosome formation occurred at the border of the triple helical DNA. In this case the tripler DNA functioned as a nucleosome barrier. We conclude that tripler DNA cannot be accommodated within a nucleosome context and thus may be used to site-specifically manipulate nucleosome organization.
引用
收藏
页码:2184 / 2191
页数:8
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