The presence of an RNA:DNA hybrid that is prone to slippage promotes termination by T7 RNA polymerase

Vadim Molodtsov, Michael Anikin, William T. McAllister

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Intrinsic termination signals for multisubunit bacterial RNA polymerases (RNAPs) encode a GC-rich stem-loop structure followed by a polyuridine [poly(U)] tract, and it has been proposed that steric clash of the stem-loop with the exit pore of the RNAP imposes a shearing force on the RNA in the downstream RNA:DNA hybrid, resulting in misalignment of the active site. The structurally unrelated T7 RNAP terminates at a similar type of signal (TΦ), suggesting a common mechanism for termination. In the absence of a hairpin (passive conditions), T7 RNAP slips efficiently in both homopolymeric A and U tracts, and we have found that replacement of the U tract in TΦ with a slippage-prone A tract still allows efficient termination. Under passive conditions, incorporation of a single G residue following a poly(U) tract (which is the situation during termination at TΦ) results in a "locked" complex that is unable to extend the transcript. Our results support a model in which transmission of the shearing force generated by steric clash of the hairpin with the exit pore is promoted by the presence of a slippery tracts downstream, resulting in alterations in the active site and the formation of a locked complex that represents an early step in the termination pathway.

Original languageEnglish (US)
Pages (from-to)3095-3107
Number of pages13
JournalJournal of Molecular Biology
Volume426
Issue number18
DOIs
StatePublished - Sep 9 2014

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DNA-Directed RNA Polymerases
Catalytic Domain
Bacterial RNA
RNA
DNA
bacteriophage T7 RNA polymerase

All Science Journal Classification (ASJC) codes

  • Molecular Biology

Cite this

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The presence of an RNA:DNA hybrid that is prone to slippage promotes termination by T7 RNA polymerase. / Molodtsov, Vadim; Anikin, Michael; McAllister, William T.

In: Journal of Molecular Biology, Vol. 426, No. 18, 09.09.2014, p. 3095-3107.

Research output: Contribution to journalArticle

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