Chemical Inhibition of Pre-mRNA Splicing in Living Saccharomyces cerevisiae

Cell Chem Biol. 2019 Mar 21;26(3):443-448.e3. doi: 10.1016/j.chembiol.2018.11.008. Epub 2019 Jan 10.

Abstract

The spliceosome mediates precursor mRNA splicing in eukaryotes, including the model organism Saccharomyces cerevisiae (yeast). Despite decades of study, no chemical inhibitors of yeast splicing in vivo are available. We have developed a system to efficiently inhibit splicing and block proliferation in living yeast cells using compounds that target the human spliceosome protein SF3B1. Potent inhibition is observed in yeast expressing a chimeric protein containing portions of human SF3B1. However, only a single point mutation in the yeast homolog of SF3B1 is needed for selective inhibition of splicing by pladienolide B, herboxidiene, or meayamycin in liquid culture. Mutations that enable inhibition also improve splicing of branch sites containing mismatches between the intron and small nuclear RNA-suggesting a link between inhibitor sensitivity and usage of weak branch sites in humans. This approach provides powerful new tools for manipulating splicing in live yeast and studies of spliceosome inhibitors.

Keywords: RNA; SF3B1; herboxidiene; inhibitor; meayamycin; pladienolide B; pre-mRNA; spliceosome; splicing; yeast.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Epoxy Compounds / chemistry
  • Epoxy Compounds / pharmacology
  • Fatty Alcohols / chemistry
  • Fatty Alcohols / pharmacology
  • Humans
  • Macrolides / chemistry
  • Macrolides / pharmacology
  • Mutagenesis
  • Phosphoproteins / chemistry
  • Phosphoproteins / metabolism
  • Pyrans / chemistry
  • Pyrans / pharmacology
  • RNA Precursors / antagonists & inhibitors
  • RNA Precursors / metabolism*
  • RNA Splicing / drug effects
  • RNA Splicing Factors / chemistry
  • RNA Splicing Factors / metabolism
  • Ribonucleoprotein, U2 Small Nuclear / genetics*
  • Ribonucleoprotein, U2 Small Nuclear / metabolism
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Sequence Alignment
  • Small Molecule Libraries / chemistry*
  • Small Molecule Libraries / metabolism
  • Small Molecule Libraries / pharmacology

Substances

  • Epoxy Compounds
  • Fatty Alcohols
  • HSH155 protein, S cerevisiae
  • Macrolides
  • Phosphoproteins
  • Pyrans
  • RNA Precursors
  • RNA Splicing Factors
  • Ribonucleoprotein, U2 Small Nuclear
  • SF3B1 protein, human
  • Saccharomyces cerevisiae Proteins
  • Small Molecule Libraries
  • meayamycin
  • pladienolide B
  • herboxidiene