title: SmD3 regulates intronic noncoding RNA biogenesis.
abstract: Accumulation of excess lipid in nonadipose tissues is associated with oxidative stress and organ dysfunction and plays an important role in diabetic complications. To elucidate molecular events critical for lipotoxicity, we used retroviral promoter trap mutagenesis to generate mutant Chinese hamster ovary cell lines resistant to lipotoxic and oxidative stress. A previous report of a mutant from this screen demonstrated that under lipotoxic conditions, small nucleolar RNAs (snoRNAs) in the rpL13a gene accumulate in the cytosol and serve as critical mediators of lipotoxic cell death. We now report a novel, independent mutant in which a single provirus disrupted one allele of the gene encoding the spliceosomal protein SmD3, creating a model of haploinsufficiency. We show that snoRNA expression and the abundance of snoRNA-containing intron lariats are decreased in SmD3 mutant cells, even though haploinsufficiency of SmD3 supports pre-mRNA splicing. The mechanism through which SmD3 regulates the expression of intronic snoRNAs likely involves effects of SmD3 on the levels of small nuclear RNAs (snRNAs) U4 and U5. Our data implicate SmD3 as a critical determinant in the processing of intronic noncoding RNAs in general and as an upstream mediator of metabolic stress response pathways through the regulation of snoRNA expression.
authors.affiliation: Diabetic Cardiovascular Disease Center and Department of Medicine, Washington University, St. Louis, Missouri, USA.
authors.forename: Benjamin S
authors.initials: BS
authors.lastname: Scruggs
authors.forename: Carlos I
authors.initials: CI
authors.lastname: Michel
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Ory
authors.forename: Jean E
authors.initials: JE
authors.lastname: Schaffer
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D057895:Haploinsufficiency; D007438:Introns; D011533:Proviruses / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics; D022661:RNA, Untranslated / Q000096:biosynthesis*; D017414:Ribonucleoprotein, U4-U6 Small Nuclear / Q000378:metabolism; D017415:Ribonucleoprotein, U5 Small Nuclear / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000235:genetics / Q000378:metabolism*; D017381:Spliceosomes / Q000235:genetics / Q000378:metabolism
chemical_list: D022661:RNA, Untranslated; D017414:Ribonucleoprotein, U4-U6 Small Nuclear; D017415:Ribonucleoprotein, U5 Small Nuclear; D017411:Ribonucleoproteins, Small Nuclear; C090866:Sm protein D3