The large N-terminal region of the Brr2 RNA helicase guides productive spliceosome activation.

Abstract:

:The Brr2 helicase provides the key remodeling activity for spliceosome catalytic activation, during which it disrupts the U4/U6 di-snRNP (small nuclear RNA protein), and its activity has to be tightly regulated. Brr2 exhibits an unusual architecture, including an ∼ 500-residue N-terminal region, whose functions and molecular mechanisms are presently unknown, followed by a tandem array of structurally similar helicase units (cassettes), only the first of which is catalytically active. Here, we show by crystal structure analysis of full-length Brr2 in complex with a regulatory Jab1/MPN domain of the Prp8 protein and by cross-linking/mass spectrometry of isolated Brr2 that the Brr2 N-terminal region encompasses two folded domains and adjacent linear elements that clamp and interconnect the helicase cassettes. Stepwise N-terminal truncations led to yeast growth and splicing defects, reduced Brr2 association with U4/U6•U5 tri-snRNPs, and increased ATP-dependent disruption of the tri-snRNP, yielding U4/U6 di-snRNP and U5 snRNP. Trends in the RNA-binding, ATPase, and helicase activities of the Brr2 truncation variants are fully rationalized by the crystal structure, demonstrating that the N-terminal region autoinhibits Brr2 via substrate competition and conformational clamping. Our results reveal molecular mechanisms that prevent premature and unproductive tri-snRNP disruption and suggest novel principles of Brr2-dependent splicing regulation.

journal_name

Genes Dev

journal_title

Genes & development

authors

Absmeier E,Wollenhaupt J,Mozaffari-Jovin S,Becke C,Lee CT,Preussner M,Heyd F,Urlaub H,Lührmann R,Santos KF,Wahl MC

doi

10.1101/gad.271528.115

subject

Has Abstract

pub_date

2015-12-15 00:00:00

pages

2576-87

issue

24

eissn

0890-9369

issn

1549-5477

pii

gad.271528.115

journal_volume

29

pub_type

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