Structure of the human activated spliceosome in three conformational states

Cell Res. 2018 Mar;28(3):307-322. doi: 10.1038/cr.2018.14. Epub 2018 Jan 23.


During each cycle of pre-mRNA splicing, the pre-catalytic spliceosome (B complex) is converted into the activated spliceosome (Bact complex), which has a well-formed active site but cannot proceed to the branching reaction. Here, we present the cryo-EM structure of the human Bact complex in three distinct conformational states. The EM map allows atomic modeling of nearly all protein components of the U2 small nuclear ribonucleoprotein (snRNP), including three of the SF3a complex and seven of the SF3b complex. The structure of the human Bact complex contains 52 proteins, U2, U5, and U6 small nuclear RNA (snRNA), and a pre-mRNA. Three distinct conformations have been captured, representing the early, mature, and late states of the human Bact complex. These complexes differ in the orientation of the Switch loop of Prp8, the splicing factors RNF113A and NY-CO-10, and most components of the NineTeen complex (NTC) and the NTC-related complex. Analysis of these three complexes and comparison with the B and C complexes reveal an ordered flux of components in the B-to-Bact and the Bact-to-B* transitions, which ultimately prime the active site for the branching reaction.

MeSH terms

  • Catalytic Domain
  • Cryoelectron Microscopy / methods
  • Humans
  • Models, Molecular*
  • Protein Conformation
  • RNA Precursors / chemistry
  • RNA, Small Nuclear / chemistry
  • Ribonucleoprotein, U2 Small Nuclear / chemistry
  • Ribonucleoprotein, U4-U6 Small Nuclear / chemistry
  • Ribonucleoprotein, U5 Small Nuclear / chemistry
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / metabolism
  • Spliceosomes / chemistry*
  • Spliceosomes / metabolism*


  • RNA Precursors
  • RNA, Small Nuclear
  • Ribonucleoprotein, U2 Small Nuclear
  • Ribonucleoprotein, U4-U6 Small Nuclear
  • Ribonucleoprotein, U5 Small Nuclear
  • Saccharomyces cerevisiae Proteins