Abstract:
:We have studied the conditions needed for the formation of stable ternary complexes by Escherichia coli RNA polymerase using a procedure in which elongation by the majority of active enzyme molecules is halted at a specific template base. Stable complexes of this sort, containing RNA chains as short as 15 nucleotides, have been formed from three different promoter sites (T7 A1, lambda PL, and E. coli rrnB P1) using di- and trinucleotides as primers in reactions limited by the presence of only three of the nucleoside triphosphate substrates. The resulting ternary complexes can be stored for at least five days without loss in activity, and provide useful reagents and substrates for studies of the properties of RNA polymerases engaged in chain elongation and termination. At all three promoter sites abortive initiation, leading to synthesis and release of oligomers up to ten nucleotides, competes with productive initiation, leading to the formation of stable elongating complexes. Thus the relative instability of ternary RNA polymerase complexes bearing transcripts shorter than ten nucleotides may be a general feature of the transcription initiation reaction.
journal_name
J Mol Bioljournal_title
Journal of molecular biologyauthors
Levin JR,Krummel B,Chamberlin MJdoi
10.1016/0022-2836(87)90512-2subject
Has Abstractpub_date
1987-07-05 00:00:00pages
85-100issue
1eissn
0022-2836issn
1089-8638pii
0022-2836(87)90512-2journal_volume
196pub_type
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