Translational control of SEPT9 isoforms is perturbed in disease.
A common feature of the mammalian septin gene family is complex genomic architecture with multiple alternate splice variants. Septin 9 has 18 distinct transcripts encoding 15 polypeptides, with two transcripts (SEPT9_v4 and v4*) encoding the same polypeptide. We have previously reported that the ratio of these distinct transcripts is altered in neoplasia, with the v4 transcript being the usual form in normal cells but v4* becoming predominant in tumours. This led us to ask what the functional differences between these two transcripts might be. The 5'-UTRs of v4 and v4* have distinct 5' ends encoded by exons 1beta (v4) and 1zeta and 2 (v4*) and a common 3' region and initiating ATG encoded within exon 3. Here we show that the two mRNAs are translated with different efficiencies and that cellular stress can alter this. A putative internal ribosome entry site can be identified in the common region of the v4 and v4* 5'-UTRs and translation is modulated by an upstream open-reading frame in the unique region of the v4 5'-UTR. Germline mutations in hereditary neuralgic amyotrophy (HNA) map to the region which is common to the two UTRs. These mutations dramatically enhance the translational efficiency of the v4 5'-UTR, leading to elevated SEPT9_v4 protein under hypoxic conditions. Our data provide a mechanistic insight into how the HNA mutations can alter the fine control of SEPT9_v4 protein and its regulation under physiologically relevant conditions and are consistent with the episodic and stress-induced nature of the clinical features of HNA.
McDade
Simon S
SS
The Centre for Cancer Research and Cell Biology, School of Medicine and Dentistry, Queen's University Belfast, Belfast City Hospital, Belfast, UK.
Hall
Peter A
PA
Russell
S E Hilary
SE
D020121:5' Untranslated Regions; D017398:Alternative Splicing; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D020968:Brachial Plexus Neuritis / Q000235:genetics* / Q000473:pathology; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D005091:Exons / Q000235:genetics; D020558:GTP Phosphohydrolases / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D014176:Protein Biosynthesis / Q000235:genetics*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D058112:Septins; D014158:Transcription, Genetic
D020121:5' Untranslated Regions; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D008156:Luciferases; D020558:GTP Phosphohydrolases; C474315:SEPTIN9 protein, human; D058112:Septins