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TitleStructure of a transcribing RNA polymerase II-U1 snRNP complex.
Journal, issue, pagesScience, Vol. 371, Issue 6526, Page 305-309, Year 2021
Publish dateJan 15, 2021
AuthorsSuyang Zhang / Shintaro Aibara / Seychelle M Vos / Dmitry E Agafonov / Reinhard Lührmann / Patrick Cramer /
PubMed AbstractTo initiate cotranscriptional splicing, RNA polymerase II (Pol II) recruits the U1 small nuclear ribonucleoprotein particle (U1 snRNP) to nascent precursor messenger RNA (pre-mRNA). Here, we report ...To initiate cotranscriptional splicing, RNA polymerase II (Pol II) recruits the U1 small nuclear ribonucleoprotein particle (U1 snRNP) to nascent precursor messenger RNA (pre-mRNA). Here, we report the cryo-electron microscopy structure of a mammalian transcribing Pol II-U1 snRNP complex. The structure reveals that Pol II and U1 snRNP interact directly. This interaction positions the pre-mRNA 5' splice site near the RNA exit site of Pol II. Extension of pre-mRNA retains the 5' splice site, leading to the formation of a "growing intron loop." Loop formation may facilitate scanning of nascent pre-mRNA for the 3' splice site, functional pairing of distant intron ends, and prespliceosome assembly. Our results provide a starting point for a mechanistic analysis of cotranscriptional spliceosome assembly and the biogenesis of mRNA isoforms by alternative splicing.
External linksScience / PubMed:33446560
MethodsEM (single particle)
Resolution3.2 - 5.9 Å
Structure data

EMDB-11972: Overall map of a transcribing RNA polymerase II-U1 snRNP complex with 145-nt RNA
PDB-7b0y: Structure of a transcribing RNA polymerase II-U1 snRNP complex
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-11973:
Focused refined map at the interface of the transcribing Pol II-U1 snRNP complex with 145-nt RNA
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-11974:
Focused refined map of U1 snRNP of the transcribing Pol II-U1 snRNP complex with 145-nt RNA
Method: EM (single particle) / Resolution: 5.9 Å

EMDB-11975:
Cryo-EM map of the transcribing Pol II-U1 snRNP complex with 155-nt RNA
Method: EM (single particle) / Resolution: 3.9 Å

Chemicals

ChemComp-ZN:
ZINC ION / Zinc

ChemComp-MG:
MAGNESIUM ION / Magnesium

Source
  • sus scrofa domesticus (domestic pig)
  • synthetic construct (others)
  • homo sapiens (human)
  • Human (human)
KeywordsAlternative Splicing / Animals / Cryoelectron Microscopy / Humans / Introns / Nucleic Acid Conformation / Protein Binding / Protein Domains / RNA Polymerase II / RNA Precursors / RNA, Messenger / Ribonucleoprotein, U1 Small Nuclear / Spliceosomes / Sus scrofa / Transcription, Genetic / SPLICING / Transcription / co-transcriptional spliceosome assembly

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