Nucleic Acids Research, Vol 24, Issue 20 3887-3895, Copyright © 1996 by Oxford University Press
JK Eadara, KG Hadlock and LC Lutter
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 BglI, SphI, KpnI and MspI restriction enzyme sites within this
region. Thus 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
trans-acting 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.
ARTICLES
Chromatin structure and factor site occupancies in an in vivo-assembled transcription elongation complex
Molecular Biology Research Program, Henry Ford Hospital, Detroit, MI 48202, USA.
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