RNA Components of Spliceosome Active Site

S. cerevisiae- 4 RNAs-~80 proteins

66% RNA

H. sapiens-5 RNAs (~400 nt)-80-100 proteins5% RNA

Complex RNA Protein Assemblies

S. cerevisiae- 4 RNAs-~80 proteins

66% RNA

H. sapiens-5 RNAs (~400 nt)-80-100 proteins5% RNA

Complex RNA Protein Assemblies

Spliceosome Proteomics

U1-70kD SNP1U1 A MUD1U1 C YHC1

U1 snRNP (3)

U2 snRNP (10)SAP155 HSH155SAP145 CUS1SAP130 RSE1SAP114 PRP21SAP62 PRP11SAP61 PRP9SAP49 HSH49U2 A' LEA1U2 B" MSL1p14 SNU17

U5 snRNP (8)U5-220kD PRP8U5-200kD BRR2U5-116kD SNU114U5-102kD PRP6U5-100kD PRP28U5-52kD SNU40U5-40kDU5-15kD DIB1

U4/U6-90kD PRP3U4/U6-60kD PRP4U4/U6-61kD PRP31U4/U6-20kD CPH1U4/U6-15.5kD SNU13

U4/U6 snRNP (5)

U4/U6.U5 tri-snRNP (3)Tri-snRNP 110kDSNU66Tri-snRNP 65kD SAD1Tri-snRNP 27kD

Sm/LSm core (33)Sm B/B' SMB1Sm D1 SMD1Sm D2 SMD2Sm D3 SMD3Sm E SME1Sm F SMX3Sm G SMX2hLSm2 LSM2hLSm3 LSM3hLSm4 LSM4hLSm6 LSM6hLSm7 LSM7

(~60 proteins)

Spliceosome+ Proteomics

SR (10)SRp75p54/SFRS1SRp55SRp40ASF/SF29G8SC35SRp30chTra2SRp20

SRm300hPrp5 PRP5hCrn CLF1CDC5L CEF1hCBP80 CBP80SF1 BBPSKIP PRP45PUF60PLRG1 PRP46hPrp19 PRP19U2AF65 MUD2U2AF35SPF30hCBP20 CBP20hFBP11 PRP40

Non-snRNP assembly (15)hPrp16 PRP16hPrp22 PRP22hPrp43 PRP43hSlu7 SLU7hPrp17 PRP17hPrp18 PRP18

Catalytic step II/late (6)

(~20-30+ proteins)

Spliceosomal RearrangementsSpliceosomal Rearrangements

GU AGA

Spliceosomal RearrangementsSpliceosomal Rearrangements

GU AGA

Spliceosomal RearrangementsSpliceosomal Rearrangements

BBP U2AFGU AGA

Spliceosomal RearrangementsSpliceosomal Rearrangements

GU AGA

Spliceosomal RearrangementsSpliceosomal Rearrangements

GU AGA

UGCUG CAUGAUG

A

U2 snRNA

Spliceosomal RearrangementsSpliceosomal Rearrangements

GU AGA

Spliceosomal Rearrangements

GUAGA

RRM•RNA Recognition

Cell 98, 835 (1999).

U2AF65•Nucleic Acid Complex

Mol Cell. 2006 23 49-59

Branch Recognition • 1(SF1•ssRNA)

Science (2001) 294, 1098-1102

Branch Recognition • 2(SF1•ssRNA)

Proc Natl Acad Sci USA. 2006 103, 1266-71.

Surface Pocket SF3b14•SF3b155

1475 1588

3.2

986 1078

1669 2034

IV

1894 -1898

RRM MPN

3' SS 5' SS

U5 snRNP Component PRP8

1 2335

U5 snRNP Component PRP8

Nat Struct Mol Biol. 2008 15 1199-205.

U5 snRNP Component PRP8

U1 snRNP

Nature. 2009 458 475-80.

U1 snRNA

5’ Splice Site Recognition