ATPγS stalls splicing after B complex formation but prior to spliceosome activation

  title={ATP$\gamma$S stalls splicing after B complex formation but prior to spliceosome activation},
  author={Dmitry E. Agafonov and Maria A van Santen and Berthold Kastner and Prakash Dube and Cindy L Will and Henning Urlaub and Reinhard L{\"u}hrmann},
  pages={1329 - 1337}
The ATP analog ATPγS inhibits pre-mRNA splicing in vitro, but there have been conflicting reports as to which step of splicing is inhibited by this small molecule and its inhibitory mechanism remains unclear. Here we have dissected the effect of ATPγS on pre-mRNA splicing in vitro. Addition of ATPγS to splicing extracts depleted of ATP inhibited both catalytic steps of splicing. At ATPγS concentrations ≥0.5 mM, precatalytic B complexes accumulate, demonstrating a block prior to or during the… 

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  • S. KimR. Lin
  • Biology, Chemistry
    Molecular and cellular biology
  • 1996
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A systematic analysis of the temporal and functional relationships among the H, E, A, and B splicing complexes concluded that E complex commits pre-mRNA to the splicing pathway and that this step is critical in determining the efficiency of mammalian spliceosome assembly.

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It appears that pre‐mRNA splicing, in common with other biological processes, can be regulated both positively and negatively by reversible protein phosphorylation.

Both phosphorylation and dephosphorylation of ASF/SF2 are required for pre-mRNA splicing in vitro.

Data is presented that suggests for the first time a requirement for SR protein dephosphorylation in pre-messenger RNA splicing in vitro and shows that deph phosphorylation of ASF/SF2 is required for thefirst transesterification reaction once the spliceosome has assembled.

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