title: Mammalian tissues defective in nonsense-mediated mRNA decay display highly aberrant splicing patterns.
abstract: BACKGROUND
  Nonsense-mediated mRNA decay (NMD) affects the outcome of alternative splicing by degrading mRNA isoforms with premature termination codons. Splicing regulators constitute important NMD targets; however, the extent to which loss of NMD causes extensive deregulation of alternative splicing has not previously been assayed in a global, unbiased manner. Here, we combine mouse genetics and RNA-seq to provide the first in vivo analysis of the global impact of NMD on splicing patterns in two primary mouse tissues ablated for the NMD factor UPF2.
  
  
  RESULTS
  We developed a bioinformatic pipeline that maps RNA-seq data to a combinatorial exon database, predicts NMD-susceptibility for mRNA isoforms and calculates the distribution of major splice isoform classes. We present a catalog of NMD-regulated alternative splicing events, showing that isoforms of 30% of all expressed genes are upregulated in NMD-deficient cells and that NMD targets all major splicing classes. Importantly, NMD-dependent effects are not restricted to premature termination codon+ isoforms but also involve an abundance of splicing events that do not generate premature termination codons. Supporting their functional importance, the latter events are associated with high intronic conservation.
  
  
  CONCLUSIONS
  Our data demonstrate that NMD regulates alternative splicing outcomes through an intricate web of splicing regulators and that its loss leads to the deregulation of a panoply of splicing events, providing novel insights into its role in core- and tissue-specific regulation of gene expression. Thus, our study extends the importance of NMD from an mRNA quality pathway to a regulator of several layers of gene expression.
authors.affiliation: The Finsen Laboratory, Rigshospitalet, Faculty of Health Sciences, University of Copenhagen, DK2200 Copenhagen, Denmark. bo.porse@finsenlab.dk
authors.forename: Joachim
authors.initials: J
authors.lastname: Weischenfeldt
authors.forename: Johannes
authors.initials: J
authors.lastname: Waage
authors.forename: Geng
authors.initials: G
authors.lastname: Tian
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.forename: Inge
authors.initials: I
authors.lastname: Damgaard
authors.forename: Janus Schou
authors.initials: JS
authors.lastname: Jakobsen
authors.forename: Karsten
authors.initials: K
authors.lastname: Kristiansen
authors.forename: Anders
authors.initials: A
authors.lastname: Krogh
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: Bo T
authors.initials: BT
authors.lastname: Porse
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D018389:Codon, Nonsense / Q000378:metabolism; D016678:Genome; D008297:Male; D008322:Mammals / Q000235:genetics / Q000378:metabolism; D051379:Mice; D059365:Nonsense Mediated mRNA Decay; D012333:RNA, Messenger / Q000378:metabolism*
chemical_list: D018389:Codon, Nonsense; D012333:RNA, Messenger