5dca › Citations

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

Figure 1.
Figure 2.
Figure 3.
Absmeier E, Wollenhaupt J, Mozaffari-Jovin S, Becke C, Lee CT, Preussner M, Heyd F, Urlaub H, Lührmann R, Santos KF, Wahl MC
OpenAccess logo Genes Dev 29 2576-87 (2015)
Cited: 40 times
EuropePMC logo PMID: 26637280

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.

Reviews - 5dca mentioned but not cited (2)

  1. Functions and regulation of the Brr2 RNA helicase during splicing. Absmeier E, Santos KF, Wahl MC. Cell Cycle 15 3362-3377 (2016)
  2. Genetics and biochemistry remain essential in the structural era of the spliceosome. Mayerle M, Guthrie C. Methods 125 3-9 (2017)

Articles - 5dca mentioned but not cited (4)

  1. The large N-terminal region of the Brr2 RNA helicase guides productive spliceosome activation. Absmeier E, Wollenhaupt J, Mozaffari-Jovin S, Becke C, Lee CT, Preussner M, Heyd F, Urlaub H, Lührmann R, Santos KF, Wahl MC. Genes Dev 29 2576-2587 (2015)
  2. Interplay of cis- and trans-regulatory mechanisms in the spliceosomal RNA helicase Brr2. Absmeier E, Becke C, Wollenhaupt J, Santos KF, Wahl MC. Cell Cycle 16 100-112 (2017)
  3. The HelQ human DNA repair helicase utilizes a PWI-like domain for DNA loading through interaction with RPA, triggering DNA unwinding by the HelQ helicase core. Jenkins T, Northall SJ, Ptchelkine D, Lever R, Cubbon A, Betts H, Taresco V, Cooper CDO, McHugh PJ, Soultanas P, Bolt EL. NAR Cancer 3 zcaa043 (2021)
  4. Intrinsically Disordered Protein Ntr2 Modulates the Spliceosomal RNA Helicase Brr2. Wollenhaupt J, Henning LM, Sticht J, Becke C, Freund C, Santos KF, Wahl MC. Biophys J 114 788-799 (2018)


Reviews citing this publication (8)

  1. Structural Insights into Nuclear pre-mRNA Splicing in Higher Eukaryotes. Kastner B, Will CL, Stark H, Lührmann R. Cold Spring Harb Perspect Biol 11 a032417 (2019)
  2. Mutations in spliceosomal proteins and retina degeneration. Růžičková Š, Staněk D. RNA Biol 14 544-552 (2017)
  3. The life of U6 small nuclear RNA, from cradle to grave. Didychuk AL, Butcher SE, Brow DA. RNA 24 437-460 (2018)
  4. Structural dynamics of the N-terminal domain and the Switch loop of Prp8 during spliceosome assembly and activation. Jia X, Sun C. Nucleic Acids Res 46 3833-3840 (2018)
  5. RNA and Proteins: Mutual Respect. Hall KB. F1000Res 6 345 (2017)
  6. Understanding pre-mRNA splicing through crystallography. Espinosa S, Zhang L, Li X, Zhao R. Methods 125 55-62 (2017)
  7. Spliceosome assembly and regulation: insights from analysis of highly reduced spliceosomes. Black CS, Whelan TA, Garside EL, MacMillan AM, Fast NM, Rader SD. RNA 29 531-550 (2023)
  8. Cellular functions of eukaryotic RNA helicases and their links to human diseases. Bohnsack KE, Yi S, Venus S, Jankowsky E, Bohnsack MT. Nat Rev Mol Cell Biol 24 749-769 (2023)

Articles citing this publication (26)

  1. Structure of a yeast activated spliceosome at 3.5 Å resolution. Yan C, Wan R, Bai R, Huang G, Shi Y. Science 353 904-911 (2016)
  2. Molecular architecture of the human U4/U6.U5 tri-snRNP. Agafonov DE, Kastner B, Dybkov O, Hofele RV, Liu WT, Urlaub H, Lührmann R, Stark H. Science 351 1416-1420 (2016)
  3. Molecular architecture of the Saccharomyces cerevisiae activated spliceosome. Rauhut R, Fabrizio P, Dybkov O, Hartmuth K, Pena V, Chari A, Kumar V, Lee CT, Urlaub H, Kastner B, Stark H, Lührmann R. Science 353 1399-1405 (2016)
  4. Mechanism of 5' splice site transfer for human spliceosome activation. Charenton C, Wilkinson ME, Nagai K. Science 364 362-367 (2019)
  5. An activated Q-SNARE/SM protein complex as a possible intermediate in SNARE assembly. Jakhanwal S, Lee CT, Urlaub H, Jahn R. EMBO J 36 1788-1802 (2017)
  6. SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing. Pozzi B, Bragado L, Will CL, Mammi P, Risso G, Urlaub H, Lührmann R, Srebrow A. Nucleic Acids Res 45 6729-6745 (2017)
  7. Substrate-assisted mechanism of RNP disruption by the spliceosomal Brr2 RNA helicase. Theuser M, Höbartner C, Wahl MC, Santos KF. Proc Natl Acad Sci U S A 113 7798-7803 (2016)
  8. Prp8 retinitis pigmentosa mutants cause defects in the transition between the catalytic steps of splicing. Mayerle M, Guthrie C. RNA 22 793-809 (2016)
  9. A new role for FBP21 as regulator of Brr2 helicase activity. Henning LM, Santos KF, Sticht J, Jehle S, Lee CT, Wittwer M, Urlaub H, Stelzl U, Wahl MC, Freund C. Nucleic Acids Res 45 7922-7937 (2017)
  10. Discovery of spiro[indole-3,2'-pyrrolidin]-2(1H)-one based inhibitors targeting Brr2, a core component of the U5 snRNP. Ito M, Iwatani M, Yamamoto T, Tanaka T, Kawamoto T, Morishita D, Nakanishi A, Maezaki H. Bioorg Med Chem 25 4753-4767 (2017)
  11. The interaction of DNA repair factors ASCC2 and ASCC3 is affected by somatic cancer mutations. Jia J, Absmeier E, Holton N, Pietrzyk-Brzezinska AJ, Hackert P, Bohnsack KE, Bohnsack MT, Wahl MC. Nat Commun 11 5535 (2020)
  12. Variability in clinical phenotypes of PRPF8-linked autosomal dominant retinitis pigmentosa correlates with differential PRPF8/SNRNP200 interactions. Escher P, Passarin O, Munier FL, Tran VH, Vaclavik V. Ophthalmic Genet 39 80-86 (2018)
  13. The inactive C-terminal cassette of the dual-cassette RNA helicase BRR2 both stimulates and inhibits the activity of the N-terminal helicase unit. Vester K, Santos KF, Kuropka B, Weise C, Wahl MC. J Biol Chem 295 2097-2112 (2020)
  14. Retinitis Pigmentosa Mutations in Bad Response to Refrigeration 2 (Brr2) Impair ATPase and Helicase Activity. Ledoux S, Guthrie C. J Biol Chem 291 11954-11965 (2016)
  15. A multi-factor trafficking site on the spliceosome remodeling enzyme BRR2 recruits C9ORF78 to regulate alternative splicing. Bergfort A, Preußner M, Kuropka B, Ilik İA, Hilal T, Weber G, Freund C, Aktaş T, Heyd F, Wahl MC. Nat Commun 13 1132 (2022)
  16. Large-scale ratcheting in a bacterial DEAH/RHA-type RNA helicase that modulates antibiotics susceptibility. Grass LM, Wollenhaupt J, Barthel T, Parfentev I, Urlaub H, Loll B, Klauck E, Antelmann H, Wahl MC. Proc Natl Acad Sci U S A 118 e2100370118 (2021)
  17. The intrinsically disordered TSSC4 protein acts as a helicase inhibitor, placeholder and multi-interaction coordinator during snRNP assembly and recycling. Bergfort A, Hilal T, Kuropka B, Ilik İA, Weber G, Aktaş T, Freund C, Wahl MC. Nucleic Acids Res 50 2938-2958 (2022)
  18. Phosphorylation by Prp4 kinase releases the self-inhibition of FgPrp31 in Fusarium graminearum. Gao X, Zhang J, Song C, Yuan K, Wang J, Jin Q, Xu JR. Curr Genet 64 1261-1274 (2018)
  19. An Allosteric Network for Spliceosome Activation Revealed by High-Throughput Suppressor Analysis in Saccharomyces cerevisiae. Brow DA. Genetics 212 111-124 (2019)
  20. Long-range allostery mediates cooperative adenine nucleotide binding by the Ski2-like RNA helicase Brr2. Absmeier E, Vester K, Ghane T, Burakovskiy D, Milon P, Imhof P, Rodnina MV, Santos KF, Wahl MC. J Biol Chem 297 100829 (2021)
  21. A Snu114-GTP-Prp8 module forms a relay station for efficient splicing in yeast. Jia J, Ganichkin OM, Preußner M, Absmeier E, Alings C, Loll B, Heyd F, Wahl MC. Nucleic Acids Res 48 4572-4584 (2020)
  22. A forward genetic screen in C. elegans identifies conserved residues of spliceosomal proteins PRP8 and SNRNP200/BRR2 with a role in maintaining 5' splice site identity. Cartwright-Acar CH, Osterhoudt K, Suzuki JMNGL, Gomez DR, Katzman S, Zahler AM. Nucleic Acids Res 50 11834-11857 (2022)
  23. Closing in on ATPase Activity by an RNA Helicase. Johnson SJ, Yim MK. Structure 28 143-144 (2020)
  24. Conformation-dependent ligand hot spots in the spliceosomal RNA helicase BRR2. Vester K, Metz A, Huber S, Loll B, Wahl MC. Acta Crystallogr D Struct Biol 79 304-317 (2023)
  25. Extended DNA threading through a dual-engine motor module of the activating signal co-integrator 1 complex. Jia J, Hilal T, Bohnsack KE, Chernev A, Tsao N, Bethmann J, Arumugam A, Parmely L, Holton N, Loll B, Mosammaparast N, Bohnsack MT, Urlaub H, Wahl MC. Nat Commun 14 1886 (2023)
  26. The Terminal Extensions of Dbp7 Influence Growth and 60S Ribosomal Subunit Biogenesis in Saccharomyces cerevisiae. Contreras J, Ruiz-Blanco Ó, Dominique C, Humbert O, Henry Y, Henras AK, de la Cruz J, Villalobo E. Int J Mol Sci 24 3460 (2023)


Quick links