Separable Roles for rent1/hUpf1 in Altered Splicing and Decay of Nonsense Transcripts

• Published in: Science (American Association for the Advancement of Science) Vol. 298; no. 5592; pp. 419 - 422
• Main Authors: Mendell, Joshua T., Colette M. J. ap Rhys, Dietz, Harry C.
• Format: Journal Article
• Language: English
• Published: Washington, DC American Association for the Advancement of Science 11.10.2002; The American Association for the Advancement of Science
• Subjects: Active Transport, Cell Nucleus; Alternative Splicing; Amino Acid Substitution; Biological and medical sciences; Blotting, Northern; Cell Nucleus - metabolism; Coding; Codon; Codon, Nonsense; Codons; Cytoplasm - metabolism; Equilibrative-Nucleoside Transporter 2 - genetics; Equilibrative-Nucleoside Transporter 2 - metabolism; Evidence; Exons; Fatty Acids, Unsaturated - pharmacology; Fundamental and applied biological sciences. Psychology; Gene Silencing; Genes, T-Cell Receptor beta; Genetic engineering; HeLa Cells; Humans; Inhibition; Mammals; Messenger RNA; Molecular and cellular biology; Molecular genetics; Mutation; Nonsense; Reading frames; Recombinant Fusion Proteins - metabolism; Ribonucleic acid; RNA; RNA Helicases - genetics; RNA Helicases - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA, Small Interfering - metabolism; Site Selection; Small interfering RNA; Splicing; T lymphocytes; Trans-Activators - genetics; Trans-Activators - metabolism; Transcription Factors - genetics; Transcription Factors - metabolism; Transcription. Transcription factor. Splicing. Rna processing; Transcripts (Written Records); Translation; United States; Vertebrata; Functional analysis; Mammalia; Messenger RNA; Splicing; Cell
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.1074428

The mechanism by which disruption of reading frame can influence pre-messenger RNA (pre-mRNA) processing is poorly understood. We assessed the role of factors essential for nonsense-mediated mRNA decay (NMD) in nonsense-mediated altered splicing (NAS) with the use of RNA interference (RNAi) in mammalian cells. Inhibition of rent1/hUpf1 expression abrogated both NMD and NAS of nonsense T cell receptor β transcripts. In contrast, inhibition of rent2/hUpf2 expression did not disrupt NAS despite achieving comparable stabilization of nonsense transcripts. We also demonstrate that NAS and NMD are genetically separable functions of rent1/hUpf1. Additionally, rent1/hUpf1 enters the nucleus where it may directly influence early events in mRNA biogenesis. This provides compelling evidence that NAS relies on a component of the nonsense surveillance machinery but is not an indirect consequence of NMD.