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Journal Article

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

MPS-Authors
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Mozaffari-Jovin,  S.
Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society;

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Lee,  C. T.
Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society;

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Urlaub,  H.
Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society;

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Lührmann,  R.
Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society;

Fulltext (public)

2240195.pdf
(Publisher version), 599KB

Supplementary Material (public)

2240195_Suppl.docx
(Supplementary material), 574KB

Citation

Absmeier, E., Wollenhaupt, J., Mozaffari-Jovin, S., Becke, C., Lee, C. T., Preussner, M., et al. (2015). The large N-terminal region of the Brr2 RNA helicase guides productive spliceosome activation. Genes and Development, 29(24), 2576-2587. doi:10.1101/gad.271528.115.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0029-4327-2
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 similar to 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 disnRNP 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.