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Warkocki Z, et al.  (2009) Reconstitution of both steps of Saccharomyces cerevisiae splicing with purified spliceosomal components. Nat Struct Mol Biol 16(12):1237-43

Abstract: The spliceosome is a ribonucleoprotein machine that removes introns from pre-mRNA in a two-step reaction. To investigate the catalytic steps of splicing, we established an in vitro splicing complementation system. Spliceosomes stalled before step 1 of this process were purified to near-homogeneity from a temperature-sensitive mutant of the RNA helicase Prp2, compositionally defined, and shown to catalyze efficient step 1 when supplemented with recombinant Prp2, Spp2 and Cwc25, thereby demonstrating that Cwc25 has a previously unknown role in promoting step 1. Step 2 catalysis additionally required Prp16, Slu7, Prp18 and Prp22. Our data further suggest that Prp2 facilitates catalytic activation by remodeling the spliceosome, including destabilizing the SF3a and SF3b proteins, likely exposing the branch site before step 1. Remodeling by Prp2 was confirmed by negative stain EM and image processing. This system allows future mechanistic analyses of spliceosome activation and catalysis.

Status: Published Type: Journal Article PubMed ID: 19935684

Topics addressed in this paper

Number of different genes curated to this paper: 147

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Topics Genes linked to topics (#1 - 10 )
BFR1 BRE5 BRR1 BRR2 BUD13 BUD31 CBF2 CDC40 CEF1 CIC1
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Topics Genes linked to topics (#11 - 20 )
CKB2 CLF1 CTK1 CTK2 CTK3 CUS1 CWC15 CWC2 CWC21 CWC22
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Topics Genes linked to topics (#21 - 30 )
CWC23 CWC24 CWC25 CWC27 DBP2 DIB1 ECM2 FYV6 GBP2 HIR2
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Topics Genes linked to topics (#31 - 40 )
HMO1 HRB1 HSH155 HSH49 IMD3 IMD4 IST3 ISY1 LEA1 LHP1
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Topics Genes linked to topics (#41 - 50 )
LIN1 LSM1 LSM2 LSM3 LSM4 LSM5 LSM6 LSM7 LSM8 LUC7
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Topics Genes linked to topics (#51 - 60 )
MRPL3 MSL1 MSS116 MUD1 MUD2 NAM8 NOP1 NOP56 NPL3 NTC20
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Topics Genes linked to topics (#61 - 70 )
NTR2 PAB1 PAT1 PML1 PRP11 PRP16 PRP18 PRP19 PRP2 PRP21
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Topics Genes linked to topics (#71 - 80 )
PRP22 PRP24 PRP28 PRP3 PRP31 PRP38 PRP39 PRP4 PRP40 PRP42
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Topics Genes linked to topics (#81 - 90 )
PRP43 PRP45 PRP46 PRP6 PRP8 PRP9 RDS3 RET1 RPB5 RPC40
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Topics Genes linked to topics (#91 - 100 )
RPC82 RPL4B RPO31 RPS13 RPS22A RRP5 RSC8 RSE1 RTS2 SAD1
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Topics Genes linked to topics (#101 - 110 )
SEC13 SEN1 SLU7 SMB1 SMD1 SMD2 SMD3 SME1 SMX2 SMX3
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Topics Genes linked to topics (#111 - 120 )
SNF2 SNP1 SNT309 SNU114 SNU13 SNU23 SNU56 SNU66 SNU71 SPP2
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Topics Genes linked to topics (#121 - 130 )
SPP381 SPP382 SSA1 SSA2 SSB1 SSB2 STT4 SUP35 SWI3 SYF1
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Topics Genes linked to topics (#131 - 140 )
SYF2 TIF4631 TOP2 UBP3 URB1 UTP20 VPS1 XRN1 YGR054W YHC1
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Topics Genes linked to topics (#141 - 147 )
YJR084W YJU2 YLR419W YPP1 YRA1 YRA2 YSF3
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