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  Yeast Prp2 liberates the 5' splice site and the branch site adenosine for catalysis of pre-mRNA splicing.

Bao, P., Höbartner, C., Hartmuth, K., & Lührmann, R. (2017). Yeast Prp2 liberates the 5' splice site and the branch site adenosine for catalysis of pre-mRNA splicing. RNA, 23(12), 1770-1779. doi:10.1261/rna.063115.117.

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 Creators:
Bao, P.1, Author           
Höbartner, C.2, Author           
Hartmuth, K.1, Author           
Lührmann, R.1, Author           
Affiliations:
1Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              
2Research Group of Nucleic Acid Chemistry, MPI for biophysical chemistry, Max Planck Society, ou_578605              

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Free keywords: Prp2 RNA helicase; RNA structure; activated spliceosome; catalytic activation
 Abstract: The RNA helicase Prp2 facilitates the remodelling of the spliceosomal Bact complex to the catalytically activated B* complex just before step one of splicing. As a high-resolution cryo-EM structure of the B* complex is currently lacking, the precise spliceosome remodelling events mediated by Prp2 remain poorly understood. To investigate the latter, we used chemical structure probing to compare the RNA structure of purified yeast Bact and B* complexes. Our studies reveal deviations from conventional RNA helices in the functionally important U6 snRNA internal stem loop and U2/U6 helix Ib in the activated Bact complex, and to a lesser extent in B*. Interestingly, the N7 of U6-G60 of the catalytic triad becomes accessible to DMS modification in the B* complex, suggesting that the Hoogsteen interaction with U6-A52 is destabilised in B*. Our data show that Prp2 action does not unwind double-stranded RNA, but enhances the flexibility of the first step reactants, the pre-mRNA's 5' splice site and branch site adenosine. Prp2 therefore appears to act primarily as an RNPase to achieve catalytic activation by liberating the first step reactants in preparation for catalysis of the first step of splicing.

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Language(s): eng - English
 Dates: 2017-09-012017-12
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1261/rna.063115.117
 Degree: -

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Title: RNA
Source Genre: Journal
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Pages: - Volume / Issue: 23 (12) Sequence Number: - Start / End Page: 1770 - 1779 Identifier: -