Protein phosphatase 1 can modulate alternative 5' splice site selection in a HeLa splicing extract.
Recent studies using HeLa in vitro splicing extracts have shown that changes in the relative concentrations of constitutive protein splicing factors can affect the choice between competing 5' splice sites in alternatively spliced mammalian pre-mRNAs. Here we report that treatment of a HeLa splicing extract with human protein phosphatase 1 strongly inhibits formation of mRNA spliced to the distal 5' splice site while stimulating relative use of the proximal 5' splice site. This effect is not observed if spliceosomes assemble prior to protein phosphatase 1 treatment. These data show that alternative splicing in HeLa extracts can be mediated by changes in protein modification as well as by changes in the relative concentration of splicing factors. Changes in protein phosphorylation may thus provide a rapid mechanism for cells to respond to stimuli that require an alteration in alternative splicing patterns.
Cardinali
B
B
European Molecular Biology Laboratory, Heidelberg, Germany.
Cohen
P T
PT
Lamond
A I
AI
D017398:Alternative Splicing; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D002474:Cell-Free System; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D007700:Kinetics; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D009838:Oligodeoxyribonucleotides; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D054645:Protein Phosphatase 1; D012322:RNA Precursors / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000378:metabolism; D014020:Tissue Extracts
D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D010750:Phosphoproteins; D012322:RNA Precursors; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D014020:Tissue Extracts; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1