Abstract：
:Transcriptional pausing by RNA polymerase (RNAP) plays an essential role in gene regulation. Pausing is modified by various elongation factors, including prokaryotic NusA, but the mechanisms underlying pausing and NusA function remain unclear. Alternative models for pausing invoke blockade events that precede translocation (on-pathway), enzyme backtracking (off-pathway), or isomerization to a nonbacktracked, elemental pause state (off-pathway). We employed an optical trapping assay to probe the motions of individual RNAP molecules transcribing a DNA template carrying tandem repeats encoding the his pause, subjecting these enzymes to controlled forces. NusA significantly decreased the pause-free elongation rate of RNAP while increasing the probability of entry into short- and long-lifetime pauses, in a manner equivalent to exerting a ~19 pN force opposing transcription. The effects of force and NusA on pause probabilities and lifetimes support a reaction scheme where nonbacktracked, elemental pauses branch off the elongation pathway from the pretranslocated state of RNAP.
journal_name
Mol Celljournal_title
Molecular cellauthors
Zhou J,Ha KS,La Porta A,Landick R,Block SMdoi
10.1016/j.molcel.2011.09.018subject
Has Abstractpub_date
2011-11-18 00:00:00pages
635-46issue
4eissn
1097-2765issn
1097-4164pii
S1097-2765(11)00818-5journal_volume
44pub_type
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