title: Regulation of multiple core spliceosomal proteins by alternative splicing-coupled nonsense-mediated mRNA decay.
abstract: Alternative splicing (AS) can regulate gene expression by introducing premature termination codons (PTCs) into spliced mRNA that subsequently elicit transcript degradation by the nonsense-mediated mRNA decay (NMD) pathway. However, the range of cellular functions controlled by this process and the factors required are poorly understood. By quantitative AS microarray profiling, we find that there are significant overlaps among the sets of PTC-introducing AS events affected by individual knockdown of the three core human NMD factors, Up-Frameshift 1 (UPF1), UPF2, and UPF3X/B. However, the levels of some PTC-containing splice variants are less or not detectably affected by the knockdown of UPF2 and/or UPF3X, compared with the knockdown of UPF1. The intron sequences flanking the affected alternative exons are often highly conserved, suggesting important regulatory roles for these AS events. The corresponding genes represent diverse cellular functions, and surprisingly, many encode core spliceosomal proteins and assembly factors. We further show that conserved, PTC-introducing AS events are enriched in genes that encode core spliceosomal proteins. Where tested, altering the expression levels of these core spliceosomal components affects the regulation of PTC-containing splice variants from the corresponding genes. Together, our results show that AS-coupled NMD can have different UPF factor requirements and is likely to regulate many general components of the spliceosome. The results further implicate general spliceosomal components in AS regulation.
authors.affiliation: Department of Molecular Genetics, Centre for Cellular and Biomolecular Research, 160 College Street, University of Toronto, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Arneet L
authors.initials: AL
authors.lastname: Saltzman
authors.forename: Yoon Ki
authors.initials: YK
authors.lastname: Kim
authors.forename: Qun
authors.initials: Q
authors.lastname: Pan
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Fagnani
authors.forename: Lynne E
authors.initials: LE
authors.lastname: Maquat
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Blencowe
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D018389:Codon, Nonsense / Q000235:genetics / Q000378:metabolism*; D017124:Conserved Sequence; D005091:Exons / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D020365:RNA Helicases; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D018389:Codon, Nonsense; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C468286:UPF2 protein, human; D020365:RNA Helicases; C101828:UPF1 protein, human