Chromatin structure and factor site occupancies in an in vivo-assembled transcription elongation complex

被引:14
作者
Eadara, JK [1 ]
Hadlock, KG [1 ]
Lutter, LC [1 ]
机构
[1] HENRY FORD HOSP, MOL BIOL RES PROGRAM, DETROIT, MI 48202 USA
关键词
D O I
10.1093/nar/24.20.3887
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
The chromatin structure specific to the SV40 late transcription elongation complex as well as the occupancy of several sites that bind transcription factors have been examined, These features have been determined by assessing blockage to restriction enzyme digestion, Cleavage specific to the elongation complex has been quantified using ternary complex analysis, This method involves radioactively labeling the complex by in vitro transcription followed by determining the extent of linearization by electrophoresis in an agarose gel, It was found that not only is the origin region devoid of nucleosomes, but there is also no stable factor occupancy at the BgII, SphI, KpnI and MspI restriction enzyme sites within this region. Th us these sites were cleaved to a high degree, meaning that the binding sites for a number of transcription factors, including OBP/TEF-1, TBP, DAP, as well as a proposed positioned nucleosome, are unoccupied in the native viral transcription elongation complex, The absence of these transacting factors from their respective binding sites in the elongation complex indicates that they bind only transiently possibly cycling on and off during the transcription cycle, This finding implies that various forms of transcription complex are assembled and disassembled during transcription and thus supports a 'hit-and-run' model of factor function.
引用
收藏
页码:3887 / 3895
页数:9
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