Nucl._Acids_Res.-2003-Alonso-3873-80.pdf (325.38 kB)
A Hox gene mutation that triggers Nonsense-mediated RNA decay and affects alternative splicing during Drosophila development
journal contributionposted on 2023-06-07, 20:18 authored by Claudio AlonsoClaudio Alonso, Michael Akam
Nonsense mutations are usually assumed to affect protein function by generating truncated protein products. Nonetheless, it is now clear that these mutations affect not just protein synthesis but also messenger RNA stability. The surveillance mechanism responsible for the detection and degradation of 'nonsense' RNA messages is termed nonsense-mediated RNA decay (NMD). Essential biochemical components of the NMD machinery have been defined in several species. Here we identify the Drosophila orthologue of one of these factors, Upf1, and document its expression during embryogenesis. To test whether NMD acts during Drosophila development, we make use of a mutation that introduces a stop codon into a variably spliced exon of the Hox gene Ultrabithorax (Ubx). Using real-time quantitative RT-PCR we demonstrate that Ubx transcripts containing the premature stop codon are expressed at lower levels than their wild type counterpart. Unexpectedly, we also find that the same mutation significantly increases the levels of a Ubx splicing isoform that lacks the exon containing the premature termination codon. These findings indicate that NMD is operational during Drosophila development and suggest that nonsense mutations may affect development by altering the spectrum of splicing products formed, as well as by reducing or eliminating protein synthesis.
- Published version
JournalNucleic Acids Research
Department affiliated with
- Evolution, Behaviour and Environment Publications
NotesHere I have conceived the original question - does the nonsense mediated RNA decay pathway operate duing Drosophila development- designed the experimental approach, done all the experiment, interpreted the collected data concluding that NMD indeed regulates gene expression during fly development and co-wrote the paper.
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