title: Multiple myeloma-associated hDIS3 mutations cause perturbations in cellular RNA metabolism and suggest hDIS3 PIN domain as a potential drug target.
abstract: hDIS3 is a mainly nuclear, catalytic subunit of the human exosome complex, containing exonucleolytic (RNB) and endonucleolytic (PIN) active domains. Mutations in hDIS3 have been found in ∼10% of patients with multiple myeloma (MM). Here, we show that these mutations interfere with hDIS3 exonucleolytic activity. Yeast harboring corresponding mutations in DIS3 show growth inhibition and changes in nuclear RNA metabolism typical for exosome dysfunction. Construction of a conditional DIS3 knockout in the chicken DT40 cell line revealed that DIS3 is essential for cell survival, indicating that its function cannot be replaced by other exosome-associated nucleases: hDIS3L and hRRP6. Moreover, HEK293-derived cells, in which depletion of endogenous wild-type hDIS3 was complemented with exogenously expressed MM hDIS3 mutants, proliferate at a slower rate and exhibit aberrant RNA metabolism. Importantly, MM mutations are synthetically lethal with the hDIS3 PIN domain catalytic mutation both in yeast and human cells. Since mutations in PIN domain alone have little effect on cell physiology, our results predict the hDIS3 PIN domain as a potential drug target for MM patients with hDIS3 mutations. It is an interesting example of intramolecular synthetic lethality with putative therapeutic potential in humans.
authors.affiliation: Department of Biophysics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland, Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, Pawinskiego 5a, 02-106 Warsaw, Poland and International Institute of Molecular and Cell Biology, Trojdena 4, 02-109 Warsaw, Poland.
authors.forename: Rafal
authors.initials: R
authors.lastname: Tomecki
authors.forename: Karolina
authors.initials: K
authors.lastname: Drazkowska
authors.forename: Iwo
authors.initials: I
authors.lastname: Kucinski
authors.forename: Krystian
authors.initials: K
authors.lastname: Stodus
authors.forename: Roman J
authors.initials: RJ
authors.lastname: Szczesny
authors.forename: Jakub
authors.initials: J
authors.lastname: Gruchota
authors.forename: Ewelina P
authors.initials: EP
authors.lastname: Owczarek
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kalisiak
authors.forename: Andrzej
authors.initials: A
authors.lastname: Dziembowski
mesh_terms: D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D063326:Exosome Multienzyme Ribonuclease Complex / Q000737:chemistry / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D009101:Multiple Myeloma / Q000235:genetics*; D009154:Mutation; D010641:Phenotype; D012313:RNA / Q000378:metabolism*; D020871:RNA Stability; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D012313:RNA; D063326:Exosome Multienzyme Ribonuclease Complex; C542283:DIS3 protein, S cerevisiae; C551543:DIS3 protein, human