RNA Components of Spliceosome Active Site. S. cerevisiae - 4 RNAs -~80 proteins 66% RNA H. sapiens -5 RNAs (~400 nt) -80-100 proteins 5% RNA Complex RNA.

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RNA Components of Spliceosome Active Site

S. cerevisiae - 4 RNAs -~80 proteins 66% RNA H. sapiens -5 RNAs (~400 nt) proteins 5% RNA Complex RNA Protein Assemblies

S. cerevisiae - 4 RNAs -~80 proteins 66% RNA H. sapiens -5 RNAs (~400 nt) proteins 5% RNA Complex RNA Protein Assemblies

Spliceosome Proteomics U1-70kDSNP1 U1 AMUD1 U1 CYHC1 U1 snRNP (3) U2 snRNP (10) SAP155HSH155 SAP145CUS1 SAP130RSE1 SAP114PRP21 SAP62PRP11 SAP61PRP9 SAP49HSH49 U2 A'LEA1 U2 B"MSL1 p14SNU17 U5 snRNP (8) U5-220kDPRP8 U5-200kDBRR2 U5-116kDSNU114 U5-102kDPRP6 U5-100kDPRP28 U5-52kDSNU40 U5-40kD U5-15kDDIB1 U4/U6-90kDPRP3 U4/U6-60kDPRP4 U4/U6-61kDPRP31 U4/U6-20kDCPH1 U4/U6-15.5kDSNU13 U4/U6 snRNP (5) U4/U6.U5 tri-snRNP (3) Tri-snRNP 110kDSNU66 Tri-snRNP 65kDSAD1 Tri-snRNP 27kD Sm/LSm core (33) Sm B/B'SMB1 Sm D1SMD1 Sm D2SMD2 Sm D3SMD3 Sm ESME1 Sm FSMX3 Sm GSMX2 hLSm2LSM2 hLSm3LSM3 hLSm4LSM4 hLSm6LSM6 hLSm7LSM7 (~60 proteins)

Spliceosome + Proteomics SR (10) SRp75 p54/SFRS1 SRp55 SRp40 ASF/SF2 9G8 SC35 SRp30c hTra2 SRp20 SRm300 hPrp5PRP5 hCrnCLF1 CDC5LCEF1 hCBP80CBP80 SF1BBP SKIPPRP45 PUF60 PLRG1PRP46 hPrp19PRP19 U2AF65MUD2 U2AF35 SPF30 hCBP20CBP20 hFBP11PRP40 Non-snRNP assembly (15) hPrp16PRP16 hPrp22PRP22 hPrp43PRP43 hSlu7SLU7 hPrp17PRP17 hPrp18PRP18 Catalytic step II/late (6) (~ proteins)

Spliceosomal Rearrangements GU AG A

Spliceosomal Rearrangements GU AG A

Spliceosomal Rearrangements BBPU2AF GU AG A

Spliceosomal Rearrangements GU AG A

Spliceosomal Rearrangements GU AG A UGCUG C AUGAUG A U2 snRNA

Spliceosomal Rearrangements GU AG A

Spliceosomal Rearrangements GU AG A

RRMRNA Recognition Cell 98, 835 (1999).

U2AF65Nucleic Acid Complex Mol Cell

Branch Recognition 1 (SF1ssRNA) Science (2001) 294,

Branch Recognition 2 (SF1ssRNA) Proc Natl Acad Sci USA ,

Surface Pocket SF3b14SF3b155

IV RRM MPN 3' SS5' SS U5 snRNP Component PRP

U5 snRNP Component PRP8 Nat Struct Mol Biol

U5 snRNP Component PRP8

U1 snRNP Nature

U1 snRNA

5’ Splice Site Recognition