Structures of the Catalytically Activated Yeast Spliceosome Reveal the Mechanism of Branching.

Wan, Ruixue; Bai, Rui; Yan, Chuangye; Lei, Jianlin; Shi, Yigong · Cell · 2019

basic_science · Level V

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Abstract

Pre-mRNA splicing is executed by the spliceosome. Structural characterization of the catalytically activated complex (B<sup>∗</sup>) is pivotal for understanding the branching reaction. In this study, we assembled the B<sup>∗</sup> complexes on two different pre-mRNAs from Saccharomyces cerevisiae and determined the cryo-EM structures of four distinct B<sup>∗</sup> complexes at overall resolutions of 2.9-3.8 Å. The duplex between U2 small nuclear RNA (snRNA) and the branch point sequence (BPS) is discretely away from the 5'-splice site (5'SS) in the three B<sup>∗</sup> complexes that are devoid of the step I splicing factors Yju2 and Cwc25. Recruitment of Yju2 into the active site brings the U2/BPS duplex into the vicinity of 5'SS, with the BPS nucleophile positioned 4 Å away from the catalytic metal M2. This analysis reveals the functional mechanism of Yju2 and Cwc25 in branching. These structures on different pre-mRNAs reveal substrate-specific conformations of the spliceosome in a major functional state.

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