期刊
JOURNAL OF CELL BIOLOGY
卷 162, 期 4, 页码 565-574出版社
ROCKEFELLER UNIV PRESS
DOI: 10.1083/jcb.200305013
关键词
insulator; chromatin; transcription; nucleus; retrotransposon
类别
资金
- NIGMS NIH HHS [R01 GM035463, GM 35463] Funding Source: Medline
Insulators might regulate gene expression by establishing and maintaining the organization of the chromatin fiber within the nucleus. Biochemical fractionation and in situ high salt extraction of lysed cells show that two known protein components of the gypsy insulator are present in the nuclear matrix. Using FISH with DNA probes located between two endogenous Su(Hw) binding sites, we show that the intervening DNA is arranged in a loop, with the two insulators located at the base. Mutations in insulator proteins, subjecting the cells to a brief heat shock, or destruction of the nuclear matrix lead to disruption of the loop. Insertion of an additional gypsy insulator in the center of the loop results in the formation of paired loops through the attachment of the inserted sequences to the nuclear matrix. These results suggest that the gypsy insulator might establish higher-order domains of chromatin structure and regulate nuclear organization by tethering the DNA to the nuclear matrix and creating chromatin loops.
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