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TitleStructural insights into branch site proofreading by human spliceosome.
Journal, issue, pagesNat Struct Mol Biol, Vol. 31, Issue 5, Page 835-845, Year 2024
Publish dateJan 9, 2024
AuthorsXiaofeng Zhang / Xiechao Zhan / Tong Bian / Fenghua Yang / Pan Li / Yichen Lu / Zhihan Xing / Rongyan Fan / Qiangfeng Cliff Zhang / Yigong Shi /
PubMed AbstractSelection of the pre-mRNA branch site (BS) by the U2 small nuclear ribonucleoprotein (snRNP) is crucial to prespliceosome (A complex) assembly. The RNA helicase PRP5 proofreads BS selection but the ...Selection of the pre-mRNA branch site (BS) by the U2 small nuclear ribonucleoprotein (snRNP) is crucial to prespliceosome (A complex) assembly. The RNA helicase PRP5 proofreads BS selection but the underlying mechanism remains unclear. Here we report the atomic structures of two sequential complexes leading to prespliceosome assembly: human 17S U2 snRNP and a cross-exon pre-A complex. PRP5 is anchored on 17S U2 snRNP mainly through occupation of the RNA path of SF3B1 by an acidic loop of PRP5; the helicase domain of PRP5 associates with U2 snRNA; the BS-interacting stem-loop (BSL) of U2 snRNA is shielded by TAT-SF1, unable to engage the BS. In the pre-A complex, an initial U2-BS duplex is formed; the translocated helicase domain of PRP5 stays with U2 snRNA and the acidic loop still occupies the RNA path. The pre-A conformation is specifically stabilized by the splicing factors SF1, DNAJC8 and SF3A2. Cancer-derived mutations in SF3B1 damage its association with PRP5, compromising BS proofreading. Together, these findings reveal key insights into prespliceosome assembly and BS selection or proofreading by PRP5.
External linksNat Struct Mol Biol / PubMed:38196034
MethodsEM (single particle)
Resolution2.5 - 3.0 Å
Structure data

31334

7evo

EMDB-31334: The cryo-EM map of the human 17S U2 snRNP core region
PDB-7evo: The cryo-EM structure of the human 17S U2 snRNP
Method: EM (single particle) / Resolution: 2.5 Å

32074

7vpx

EMDB-32074, PDB-7vpx:
The cryo-EM structure of the human pre-A complex
Method: EM (single particle) / Resolution: 3.0 Å

Chemicals

ChemComp-9B0:
[(2~{S},3~{S},4~{E},6~{S},7~{R},10~{R})-3,7-dimethyl-2-[(2~{E},4~{E},6~{R})-6-methyl-6-oxidanyl-7-[(2~{R},3~{R})-3-[(2~{R},3~{S})-3-oxidanylpentan-2-yl]oxiran-2-yl]hepta-2,4-dien-2-yl]-7,10-bis(oxidanyl)-12-oxidanylidene-1-oxacyclododec-4-en-6-yl] 4-cycloheptylpiperazine-1-carboxylate

ChemComp-ZN:
Unknown entry

ChemComp-SJT:
spliceostatin A (form II)

Source
  • homo sapiens (human)
  • unidentified adenovirus
KeywordsSPLICING / U2 snRNP / PRP5 / SF3B1 / human spliceosome / pre-A complex / A complex / U1 snRNP

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