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Biophys J. 2010 Oct 20;99(8):2577-86. doi: 10.1016/j.bpj.2010.08.010.

RNA polymerase II with open and closed trigger loops: active site dynamics and nucleic acid translocation.

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1
Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, USA. feig@msu.edu

Abstract

RNA polymerase II is the central eukaryotic enzyme in transcription from DNA to RNA. The dynamics of RNA polymerase II is described from molecular-dynamics simulations started from two crystal structures with open and closed trigger loop (TL) forms. Dynamic transitions between neutral and forward translocated states were observed, especially for the downstream DNA duplex. Dynamic rearrangements were also seen in the active site environment, including conformations in which the active site nucleotide assumed a possibly precatalytic conformation in close proximity to the terminal 3'-hydroxyl of the nascent RNA. Because nucleic acid translocation was observed primarily in the simulations with an open TL structure, whereas close approach of the active site nucleotide to the terminal RNA ribose predominantly occurred in the closed TL structure, a modified Brownian ratchet mechanism is proposed whereby thermally driven translocation is only possible with an open TL, and fidelity control and catalysis require TL closing.

PMID:
20959099
PMCID:
PMC2956212
DOI:
10.1016/j.bpj.2010.08.010
[Indexed for MEDLINE]
Free PMC Article
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