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Structure paper

TitleStructural basis of branch site recognition by the human spliceosome.
Journal, issue, pagesScience, Vol. 375, Issue 6576, Page 50-57, Year 2022
Publish dateJan 7, 2022
AuthorsJonas Tholen / Michal Razew / Felix Weis / Wojciech P Galej /
PubMed AbstractRecognition of the intron branch site (BS) by the U2 small nuclear ribonucleoprotein (snRNP) is a critical event during spliceosome assembly. In mammals, BS sequences are poorly conserved, and ...Recognition of the intron branch site (BS) by the U2 small nuclear ribonucleoprotein (snRNP) is a critical event during spliceosome assembly. In mammals, BS sequences are poorly conserved, and unambiguous intron recognition cannot be achieved solely through a base-pairing mechanism. We isolated human 17 U2 snRNP and reconstituted in vitro its adenosine 5´-triphosphate (ATP)–dependent remodeling and binding to the pre–messenger RNA substrate. We determined a series of high-resolution (2.0 to 2.2 angstrom) structures providing snapshots of the BS selection process. The substrate-bound U2 snRNP shows that SF3B6 stabilizes the BS:U2 snRNA duplex, which could aid binding of introns with poor sequence complementarity. ATP-dependent remodeling uncoupled from substrate binding captures U2 snRNA in a conformation that competes with BS recognition, providing a selection mechanism based on branch helix stability.
External linksScience / PubMed:34822310 / PubMed Central
MethodsEM (single particle)
Resolution2.05 - 3.3 Å
Structure data

13793

7q3l

EMDB-13793: Human 17S U2 snRNP 5' domain core
PDB-7q3l: Human 17S U2 snRNP 5' domain
Method: EM (single particle) / Resolution: 2.21 Å

EMDB-13810:
Human 17S U2 snRNP HEAT repeats
Method: EM (single particle) / Resolution: 2.9 Å

13811

7q4o

EMDB-13811, PDB-7q4o:
Substrate-bound A-like U2 snRNP
Method: EM (single particle) / Resolution: 2.1 Å

13812

7q4p

EMDB-13812, PDB-7q4p:
U2 snRNP after ATP-dependent remodelling
Method: EM (single particle) / Resolution: 2.15 Å

EMDB-13813:
A-like U2 snRNP medium-resolution map
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-13814:
AMP-PCP-treated A-like U2 snRNP
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-13815:
U2 snRNP 5' domain high resolution map
Method: EM (single particle) / Resolution: 2.05 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-HOH:
WATER

Source
  • homo sapiens (human)
  • synthetic construct (others)
KeywordsNUCLEAR PROTEIN / snRNP / Spliceosome / U2 snRNP / Splicing

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