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  Structural insights into how Prp5 proofreads the pre-mRNA branch site

Zhang, Z., Rigo, N., Dybkov, O., Fourmann, J. B., Will, C. L., Kumar, V., et al. (2021). Structural insights into how Prp5 proofreads the pre-mRNA branch site. Nature, 596(7871), 296-300. doi:10.1038/s41586-021-03789-5.

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 Creators:
Zhang, Z.1, Author              
Rigo, N.2, Author              
Dybkov, O.3, Author              
Fourmann, J. B.2, Author              
Will, C. L.3, Author              
Kumar, V.2, Author              
Urlaub, H.4, Author              
Stark, H.1, Author              
Lührmann, R.3, Author              
Affiliations:
1Department of Structural Dynamics, MPI for Biophysical Chemistry, Max Planck Society, ou_2205645              
2Department of Cellular Biochemistry, MPI for Biophysical Chemistry, Max Planck Society, ou_578576              
3Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              
4Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society, ou_578613              

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Free keywords: Cryoelectron microscopy; RNA splicing
 Abstract: During the splicing of introns from precursor messenger RNAs (pre-mRNAs), the U2 small nuclear ribonucleoprotein (snRNP) must undergo stable integration into the spliceosomal A complex-a poorly understood, multistep process that is facilitated by the DEAD-box helicase Prp5 (refs. 1-4). During this process, the U2 small nuclear RNA (snRNA) forms an RNA duplex with the pre-mRNA branch site (the U2-BS helix), which is proofread by Prp5 at this stage through an unclear mechanism5. Here, by deleting the branch-site adenosine (BS-A) or mutating the branch-site sequence of an actin pre-mRNA, we stall the assembly of spliceosomes in extracts from the yeast Saccharomyces cerevisiae directly before the A complex is formed. We then determine the three-dimensional structure of this newly identified assembly intermediate by cryo-electron microscopy. Our structure indicates that the U2-BS helix has formed in this pre-A complex, but is not yet clamped by the HEAT domain of the Hsh155 protein (Hsh155HEAT), which exhibits an open conformation. The structure further reveals a large-scale remodelling/repositioning of the U1 and U2 snRNPs during the formation of the A complex that is required to allow subsequent binding of the U4/U6.U5 tri-snRNP, but that this repositioning is blocked in the pre-A complex by the presence of Prp5. Our data suggest that binding of Hsh155HEAT to the bulged BS-A of the U2-BS helix triggers closure of Hsh155HEAT, which in turn destabilizes Prp5 binding. Thus, Prp5 proofreads the branch site indirectly, hindering spliceosome assembly if branch-site mutations prevent the remodelling of Hsh155HEAT. Our data provide structural insights into how a spliceosomal helicase enhances the fidelity of pre-mRNA splicing.

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Language(s): eng - English
 Dates: 2021-08-042021-08-12
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1038/s41586-021-03789-5
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Title: Nature
Source Genre: Journal
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Pages: - Volume / Issue: 596 (7871) Sequence Number: - Start / End Page: 296 - 300 Identifier: -