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TitleMechanism of exon ligation by human spliceosome.
Journal, issue, pagesMol Cell, Vol. 82, Issue 15, Page 2769-22778.e4, Year 2022
Publish dateAug 4, 2022
AuthorsXiechao Zhan / Yichen Lu / Xiaofeng Zhang / Chuangye Yan / Yigong Shi /
PubMed AbstractPre-mRNA splicing involves two sequential reactions: branching and exon ligation. The C complex after branching undergoes remodeling to become the C complex, which executes exon ligation. Here, we ...Pre-mRNA splicing involves two sequential reactions: branching and exon ligation. The C complex after branching undergoes remodeling to become the C complex, which executes exon ligation. Here, we report cryo-EM structures of two intermediate human spliceosomal complexes, pre-C-I and pre-C-II, both at 3.6 Å. In both structures, the 3' splice site is already docked into the active site, the ensuing 3' exon sequences are anchored on PRP8, and the step II factor FAM192A contacts the duplex between U2 snRNA and the branch site. In the transition of pre-C-I to pre-C-II, the step II factors Cactin, FAM32A, PRKRIP1, and SLU7 are recruited. Notably, the RNA helicase PRP22 is positioned quite differently in the pre-C-I, pre-C-II, and C complexes, suggesting a role in 3' exon binding and proofreading. Together with information on human C and C complexes, our studies recapitulate a molecular choreography of the C-to-C transition, revealing mechanistic insights into exon ligation.
External linksMol Cell / PubMed:35705093
MethodsEM (single particle)
Resolution3.6 - 4.3 Å
Structure data

EMDB-32317, PDB-7w59:
The cryo-EM structure of human pre-C*-I complex
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-32318: The cryo-EM map of PRP22 region of human pre-C*-I complex
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-32319, PDB-7w5a:
The cryo-EM structure of human pre-C*-II complex
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-32320: The cryo-EM map of PRP22 region of human pre-C*-II complex
Method: EM (single particle) / Resolution: 4.0 Å

EMDB-32321, PDB-7w5b:
The cryo-EM structure of human C* complex
Method: EM (single particle) / Resolution: 4.3 Å

Chemicals

ChemComp-IHP:
INOSITOL HEXAKISPHOSPHATE

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

Source
  • homo sapiens (human)
  • unidentified adenovirus
  • human (human)
KeywordsSPLICING / spliceosome / C* complex / RNA splicing / PRP22 / exon ligation / FAM192A / human spliceosome

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