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  Structural basis of transcription inhibition by alpha-amanitin and implications for RNA polymerase II translocation.

Brueckner, F., & Cramer, P. (2008). Structural basis of transcription inhibition by alpha-amanitin and implications for RNA polymerase II translocation. Nature Structural and Molecular Biology, 15, 811-818. doi:10.1038/nsmb.1458.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0015-7C12-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-8327-B
Genre: Journal Article

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Brueckner, F., Author
Cramer, P.1, Author              
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1Department of Molecular Biology, MPI for Biophysical Chemistry, Max Planck Society, ou_1863498              

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 Abstract: To study how RNA polymerase II translocates after nucleotide incorporation, we prepared elongation complex crystals in which pre- and post-translocation states interconvert. Crystal soaking with the inhibitor alpha-amanitin locked the elongation complex in a new state, which was refined at 3.4-Å resolution and identified as a possible translocation intermediate. The DNA base entering the active site occupies a 'pretemplating' position above the central bridge helix, which is shifted and occludes the templating position. A leucine residue in the trigger loop forms a wedge at the shifted bridge helix, but moves by 13 Å to close the active site during nucleotide incorporation. Our results support a Brownian ratchet mechanism that involves swinging of the trigger loop between open, wedged and closed positions, and suggest that alpha-amanitin impairs nucleotide incorporation and translocation by trapping the trigger loop and bridge helix.

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Language(s): eng - English
 Dates: 2008-06-132008
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1038/nsmb.1458
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Title: Nature Structural and Molecular Biology
Source Genre: Journal
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Pages: - Volume / Issue: 15 Sequence Number: - Start / End Page: 811 - 818 Identifier: -