National Natural Science Foundation of China (NSFC)
31930059
中国
引用
ジャーナル: Cell Res / 年: 2024 タイトル: Structural insights into human exon-defined spliceosome prior to activation. 著者: Wenyu Zhang / Xiaofeng Zhang / Xiechao Zhan / Rui Bai / Jianlin Lei / Chuangye Yan / Yigong Shi / 要旨: Spliceosome is often assembled across an exon and undergoes rearrangement to span a neighboring intron. Most states of the intron-defined spliceosome have been structurally characterized. However, ...Spliceosome is often assembled across an exon and undergoes rearrangement to span a neighboring intron. Most states of the intron-defined spliceosome have been structurally characterized. However, the structure of a fully assembled exon-defined spliceosome remains at large. During spliceosome assembly, the pre-catalytic state (B complex) is converted from its precursor (pre-B complex). Here we report atomic structures of the exon-defined human spliceosome in four sequential states: mature pre-B, late pre-B, early B, and mature B. In the previously unknown late pre-B state, U1 snRNP is already released but the remaining proteins are still in the pre-B state; unexpectedly, the RNAs are in the B state, with U6 snRNA forming a duplex with 5'-splice site and U5 snRNA recognizing the 3'-end of the exon. In the early and mature B complexes, the B-specific factors are stepwise recruited and specifically recognize the exon 3'-region. Our study reveals key insights into the assembly of the exon-defined spliceosomes and identifies mechanistic steps of the pre-B-to-B transition.
A: pre-mRNA 5A: U5 snRNA 5B: Pre-mRNA-processing-splicing factor 8 5C: 116 kDa U5 small nuclear ribonucleoprotein component 5D: U5 small nuclear ribonucleoprotein 200 kDa helicase 5E: U5 small nuclear ribonucleoprotein 40 kDa protein 5a: Isoform SM-B of Small nuclear ribonucleoprotein-associated proteins B and B' 5b: Small nuclear ribonucleoprotein Sm D1 5c: Small nuclear ribonucleoprotein Sm D2 5d: Small nuclear ribonucleoprotein F 5e: Small nuclear ribonucleoprotein E 5f: Small nuclear ribonucleoprotein G 5g: Small nuclear ribonucleoprotein Sm D3 6A: U6 snRNA 6a: U6 snRNA-associated Sm-like protein LSm2 6b: U6 snRNA-associated Sm-like protein LSm3 6c: U6 snRNA-associated Sm-like protein LSm4 6d: U6 snRNA-associated Sm-like protein LSm5 6e: U6 snRNA-associated Sm-like protein LSm6 6f: U6 snRNA-associated Sm-like protein LSm7 6g: U6 snRNA-associated Sm-like protein LSm8 4A: U4 snRNA 4B: U4/U6 small nuclear ribonucleoprotein Prp3 4C: U4/U6 small nuclear ribonucleoprotein Prp4 4D: U4/U6 small nuclear ribonucleoprotein Prp31 4E: NHP2-like protein 1 4F: Thioredoxin-like protein 4A 4G: Pre-mRNA-processing factor 6 4H: Peptidyl-prolyl cis-trans isomerase H 4I: WW domain-binding protein 4 4J: U4/U6.U5 tri-snRNP-associated protein 1 4Z: WD40 repeat-containing protein SMU1 4a: Isoform SM-B of Small nuclear ribonucleoprotein-associated proteins B and B' 4b: Small nuclear ribonucleoprotein Sm D1 4c: Small nuclear ribonucleoprotein Sm D2 4d: Small nuclear ribonucleoprotein F 4e: Small nuclear ribonucleoprotein E 4f: Small nuclear ribonucleoprotein G 4g: Small nuclear ribonucleoprotein Sm D3 2A: U2 snRNA 2B: U2 small nuclear ribonucleoprotein A' 2C: U2 small nuclear ribonucleoprotein B'' 2D: Splicing factor 3A subunit 1 2E: Splicing factor 3A subunit 2 2F: Splicing factor 3A subunit 3 2G: Splicing factor 3B subunit 1 2H: Splicing factor 3B subunit 2 2I: Splicing factor 3B subunit 3 2J: Splicing factor 3B subunit 4 2K: Splicing factor 3B subunit 6 2L: PHD finger-like domain-containing protein 5A 2M: Splicing factor 3B subunit 5 2a: Isoform SM-B of Small nuclear ribonucleoprotein-associated proteins B and B' 2b: Small nuclear ribonucleoprotein Sm D1 2c: Small nuclear ribonucleoprotein Sm D2 2d: Small nuclear ribonucleoprotein F 2e: Small nuclear ribonucleoprotein E 2f: Small nuclear ribonucleoprotein G 2g: Small nuclear ribonucleoprotein Sm D3 ヘテロ分子
分子量: 273974.250 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: Q6P2Q9
#4: タンパク質
116kDaU5smallnuclearribonucleoproteincomponent / Elongation factor Tu GTP-binding domain-containing protein 2 / SNU114 homolog / hSNU114 / U5 snRNP- ...Elongation factor Tu GTP-binding domain-containing protein 2 / SNU114 homolog / hSNU114 / U5 snRNP-specific protein / 116 kDa / U5-116 kDa
分子量: 109560.625 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: Q15029
#7: タンパク質
IsoformSM-BofSmallnuclearribonucleoprotein-associatedproteinsBandB' / snRNP-B / Sm protein B/B' / Sm-B/B' / SmB/B'
分子量: 23686.004 Da / 分子数: 3 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: P14678
#26: タンパク質
NHP2-likeprotein1 / High mobility group-like nuclear protein 2 homolog 1 / OTK27 / SNU13 homolog / hSNU13 / U4/U6.U5 ...High mobility group-like nuclear protein 2 homolog 1 / OTK27 / SNU13 homolog / hSNU13 / U4/U6.U5 small nuclear ribonucleoprotein SNU13 / U4/U6.U5 tri-snRNP 15.5 kDa protein
分子量: 14191.524 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: P55769
#27: タンパク質
Thioredoxin-likeprotein4A / DIM1 protein homolog / Spliceosomal U5 snRNP-specific 15 kDa protein / Thioredoxin-like U5 snRNP ...DIM1 protein homolog / Spliceosomal U5 snRNP-specific 15 kDa protein / Thioredoxin-like U5 snRNP protein U5-15kD
分子量: 16807.346 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: P83876
#28: タンパク質
Pre-mRNA-processingfactor6 / Androgen receptor N-terminal domain-transactivating protein 1 / ANT-1 / PRP6 homolog / U5 snRNP- ...Androgen receptor N-terminal domain-transactivating protein 1 / ANT-1 / PRP6 homolog / U5 snRNP-associated 102 kDa protein / U5-102 kDa protein
分子量: 107092.242 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: O94906
#29: タンパク質
Peptidyl-prolylcis-transisomeraseH / PPIase H / Rotamase H / Small nuclear ribonucleoprotein particle-specific cyclophilin H / CypH / U- ...PPIase H / Rotamase H / Small nuclear ribonucleoprotein particle-specific cyclophilin H / CypH / U-snRNP-associated cyclophilin SnuCyp-20 / USA-CYP
分子量: 58536.105 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: O43172
#25: タンパク質
U4/U6smallnuclearribonucleoproteinPrp31 / Pre-mRNA-processing factor 31 / Serologically defined breast cancer antigen NY-BR-99 / U4/U6 snRNP ...Pre-mRNA-processing factor 31 / Serologically defined breast cancer antigen NY-BR-99 / U4/U6 snRNP 61 kDa protein / Protein 61K / hPrp31
分子量: 55528.969 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: Q8WWY3
+
U2 small nuclear ribonucleoprotein ... , 2種, 2分子 2B2C
#34: タンパク質
U2smallnuclearribonucleoproteinA' / U2 snRNP A'
分子量: 28456.584 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: P09661
#35: タンパク質
U2smallnuclearribonucleoproteinB'' / U2 snRNP B''
分子量: 25524.367 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: P08579
+
Splicing factor 3A subunit ... , 3種, 3分子 2D2E2F
#36: タンパク質
Splicingfactor3Asubunit1 / SF3a120 / Spliceosome-associated protein 114 / SAP 114
分子量: 88991.094 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: Q15459
#37: タンパク質
Splicingfactor3Asubunit2 / SF3a66 / Spliceosome-associated protein 62 / SAP 62
分子量: 49327.355 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: Q15428
#38: タンパク質
Splicingfactor3Asubunit3 / SF3a60 / Spliceosome-associated protein 61 / SAP 61
分子量: 58934.844 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 参照: UniProt: Q12874