Nucleic Acids Research, Vol 25, Issue 13 2640-2647, Copyright © 1997 by Oxford University Press
T Kubori and N Shimamoto
Transcription initiation is accompanied with iterative synthesis and
release of short transcripts. The molar ratio of enzyme to template was
found to be critical for the amounts and distribution of the abortive
products synthesized by Escherichia coli RNA polymerase from several
promoters. At a high ratio abortive synthesis of 4-8 nt were enhanced at
thelambda P R promoter. Removing excess RNA polymerase just before
initiation, achieved by washing immobilized transcription complexes,
prevented this enhancement. At this high ratio synthesis of an unexpected 6
nt transcript was enhanced when the enzyme stalled at position +32, but not
when it stalled at position +73. This transcript had misincorporations at
its fifth and sixth positions, probably due to slippage. Hydroxyl radical
footprinting of the complex stalled at +32 in the presence of excess enzyme
showed that more than one molecule of RNA polymerase was tandemly bound to
the same DNA. These results suggest that: (i) when RNA polymerase molecules
are tandemly transcribing the same DNA, transient collisions enhance
abortive synthesis by the trailing molecule; (ii) when the leading
polymerase stalled in the initially transcribed region blocks progression
of the trailing polymerase, the latter can commit misincorporations,
probably due to slippage synthesis.
ARTICLES
Physical interference between escherichia coli RNA polymerase molecules transcribing in tandem enhances abortive synthesis and misincorporation
Structural Biology Center, National Institute of Genetics and Department of Genetics, School of Life Science, The Graduate University for Advanced Studies, Mishima, Shizuoka 411, Japan.
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