The Prp19/Pso4 core complex undergoes ubiquitylation and structural alterations in response to DNA damage.
Prp19/Pso4, a U-box containing E3 ligase, has a demonstrated role in pre-mRNA splicing, but has also been implicated in both yeast and mammalian cells as having a direct role in DNA damage processing. In this report, we provide further evidence in support of this latter assertion. We show that hPrp19 forms an ubiquitylated oligomeric species that is resistant to disruption by SDS gel electrophoresis under nonreducing conditions suggesting that is mediated by a thiolester between ubiquitin and a cysteine residue in Prp19. The level of this species is significantly enhanced upon treatment of cells with DNA damaging agents, and its association with chromatin is increased. In addition, hPrp19 is known to form a stable core complex with Cdc5L, Plrg1, and Spf27; however, ubiquitylated hPrp19 fails to interact with either Cdc5L or Plrg1 indicating that DNA damage can induce profound alterations to the hPrp19 core complex. Finally, we show that overexpression of hPrp19 in human cells provides a pro-survival affect in that it reduces the levels of apoptosis observed after exposure of cells to DNA damage.
Lu
Xiaoyan
X
Department of Cancer Genetics, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
Legerski
Randy J
RJ
D017209:Apoptosis / Q000502:physiology; D002352:Carrier Proteins / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism; D002843:Chromatin / Q000187:drug effects; D004249:DNA Damage / Q000502:physiology*; D045643:DNA Repair Enzymes; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D009687:Nuclear Proteins; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D025801:Ubiquitin / Q000378:metabolism*
C508357:CDC5L protein, human; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D002843:Chromatin; D009687:Nuclear Proteins; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins; D025801:Ubiquitin; D012967:Sodium Dodecyl Sulfate; D008741:Methyl Methanesulfonate; D045643:DNA Repair Enzymes; C417041:PRPF19 protein, human