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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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Presentation on theme: "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."— Presentation transcript:

1 RNA Components of Spliceosome Active Site

2 S. cerevisiae - 4 RNAs -~80 proteins 66% RNA H. sapiens -5 RNAs (~400 nt) -80-100 proteins 5% RNA Complex RNA Protein Assemblies

3 S. cerevisiae - 4 RNAs -~80 proteins 66% RNA H. sapiens -5 RNAs (~400 nt) -80-100 proteins 5% RNA Complex RNA Protein Assemblies

4 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)

5 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) (~20-30 + proteins)

6 Spliceosomal Rearrangements GU AG A

7 Spliceosomal Rearrangements GU AG A

8 Spliceosomal Rearrangements BBPU2AF GU AG A

9 Spliceosomal Rearrangements GU AG A

10 Spliceosomal Rearrangements GU AG A UGCUG C AUGAUG A U2 snRNA

11 Spliceosomal Rearrangements GU AG A

12 Spliceosomal Rearrangements GU AG A

13 RRMRNA Recognition Cell 98, 835 (1999).

14 U2AF65Nucleic Acid Complex Mol Cell. 2006 23 49-59

15 Branch Recognition 1 (SF1ssRNA) Science (2001) 294, 1098-1102

16 Branch Recognition 2 (SF1ssRNA) Proc Natl Acad Sci USA. 2006 103, 1266-71.

17 Surface Pocket SF3b14SF3b155

18 14751588 3.2 986 1078 16692034 IV 1894 -1898 RRM MPN 3' SS5' SS U5 snRNP Component PRP8 12335

19 U5 snRNP Component PRP8 Nat Struct Mol Biol. 2008 15 1199-205.

20 U5 snRNP Component PRP8

21 U1 snRNP Nature. 2009 458 475-80.

22 U1 snRNA

23 5’ Splice Site Recognition

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