title: Conserved regions of the DMD 3' UTR regulate translation and mRNA abundance in cultured myotubes.
abstract: Duchenne muscular dystrophy (DMD), a severe muscle-wasting disease, is caused by mutations in the DMD gene, which encodes for the protein dystrophin. Its regulation is of therapeutic interest as even small changes in expression of functional dystrophin can significantly impact the severity of DMD. While tissue-specific distribution and transcriptional regulation of several DMD mRNA isoforms has been well characterized, the post-transcriptional regulation of dystrophin synthesis is not well understood. Here, we utilize qRTPCR and a quantitative dual-luciferase reporter assay to examine the effects of isoform specific DMD 5' UTRs and the highly conserved DMD 3' UTR on mRNA abundance and translational control of gene expression in C2C12 cells. The 5' UTRs were shown to initiate translation with low efficiency in both myoblasts and myotubes. Whereas, two large highly conserved elements in the 3' UTR, which overlap the previously described Lemaire A and D regions, increase mRNA levels and enhance translation upon differentiation of myoblasts into myotubes. The results presented here implicate an important role for DMD UTRs in dystrophin expression and delineate the cis-acting elements required for the myotube-specific regulation of steady-state mRNA levels and translational enhancer activity found in the DMD 3' UTR.
authors.affiliation: Department of Human Genetics, University of Utah, Salt Lake City, UT, United States.
authors.forename: C Aaron
authors.initials: CA
authors.lastname: Larsen
authors.affiliation: Department of Human Genetics, University of Utah, Salt Lake City, UT, United States. Electronic address: mhoward@genetics.utah.edu.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Howard
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D002460:Cell Line; D017124:Conserved Sequence; D016189:Dystrophin / Q000235:genetics* / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D051379:Mice; D024510:Muscle Development / Q000235:genetics / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D032446:Myoblasts / Q000378:metabolism; D014176:Protein Biosynthesis / Q000502:physiology*; D020033:Protein Isoforms; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D020413:3' Untranslated Regions; C484258:DMD protein, human; D016189:Dystrophin; D020033:Protein Isoforms; D012333:RNA, Messenger
keywords: 3′ UTR; 5′ UTR; DMD; Duchenne muscular dystrophy; Translational control