Studitsky V M, Clark D J, Felsenfeld G
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases National Institutes of Health Bethesda, Maryland 20892.
Cell. 1994 Jan 28;76(2):371-82. doi: 10.1016/0092-8674(94)90343-3.
The mechanism by which nucleosome cores are displaced and re-formed during transcription in vitro has been investigated. A nucleosome core was assembled on a short linear DNA template (227 bp) containing an SP6 RNA polymerase promoter and a nucleosome-positioning sequence. Transcription induced the translocation of the nucleosome core over 75 or 80 bp to two positions at the other end of the template, blocking the promoter. At low rNTP concentrations, transfer occurred only on the same template molecule, even in the presence of large excesses of competitor DNA. On a longer template (262 bp), nucleosome core position after transcription depended on its position before transcription. The data suggest that the octamer transfers without dissociation from DNA and provide strong evidence for a translocation mechanism in which DNA ahead of the polymerase uncoils from the octamer as the DNA behind coils around it. In this way, the octamer steps around the transcribing polymerase.
体外转录过程中核小体核心被置换和重新形成的机制已被研究。在一个包含SP6 RNA聚合酶启动子和核小体定位序列的短线性DNA模板(227 bp)上组装了一个核小体核心。转录诱导核小体核心在模板上移动75或80 bp至模板另一端的两个位置,从而阻断启动子。在低rNTP浓度下,即使存在大量过量的竞争DNA,转移也仅发生在同一模板分子上。在更长的模板(262 bp)上,转录后核小体核心的位置取决于其转录前的位置。数据表明八聚体在不与DNA解离的情况下转移,并为一种易位机制提供了有力证据,即随着聚合酶后面的DNA围绕其缠绕,聚合酶前面的DNA从八聚体上解旋。通过这种方式,八聚体围绕转录中的聚合酶移动。