title: Novel exons in the tbx5 gene locus generate protein isoforms with distinct expression domains and function.
abstract: TBX5 is the gene mutated in Holt-Oram syndrome, an autosomal dominant disorder with complex heart and limb deformities. Its protein product is a member of the T-box family of transcription factors and an evolutionarily conserved dosage-sensitive regulator of heart and limb development. Understanding TBX5 regulation is therefore of paramount importance. Here we uncover the existence of novel exons and provide evidence that TBX5 activity may be extensively regulated through alternative splicing to produce protein isoforms with differing N- and C-terminal domains. These isoforms are also present in human heart, indicative of an evolutionarily conserved regulatory mechanism. The newly identified isoforms have different transcriptional properties and can antagonize TBX5a target gene activation. Droplet Digital PCR as well as immunohistochemistry with isoform-specific antibodies reveal differential as well as overlapping expression domains. In particular, we find that the predominant isoform in skeletal myoblasts is Tbx5c, and we show that it is dramatically up-regulated in differentiating myotubes and is essential for myotube formation. Mechanistically, TBX5c antagonizes TBX5a activation of pro-proliferative signals such as IGF-1, FGF-10, and BMP4. The results provide new insight into Tbx5 regulation and function that will further our understanding of its role in health and disease. The finding of new exons in the Tbx5 locus may also be relevant to mutational screening especially in the 30% of Holt-Oram syndrome patients with no mutations in the known TBX5a exons.
authors.affiliation: From the Laboratory of Molecular Genetics and Cardiac Regeneration, Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Ontario K1N 6N5 and.
authors.forename: Abir
authors.initials: A
authors.lastname: Yamak
authors.affiliation: the Graduate Program in Molecular Biology, Institut de Recherches Cliniques de Montréal (IRCM), Université de Montréal, Montréal, Québec H2W 1R7, Canada.
authors.forename: Romain O
authors.initials: RO
authors.lastname: Georges
authors.affiliation: From the Laboratory of Molecular Genetics and Cardiac Regeneration, Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Ontario K1N 6N5 and.
authors.forename: Massomeh
authors.initials: M
authors.lastname: Sheikh-Hassani
authors.affiliation: the Graduate Program in Molecular Biology, Institut de Recherches Cliniques de Montréal (IRCM), Université de Montréal, Montréal, Québec H2W 1R7, Canada.
authors.forename: Martin
authors.initials: M
authors.lastname: Morin
authors.affiliation: From the Laboratory of Molecular Genetics and Cardiac Regeneration, Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Ontario K1N 6N5 and.
authors.forename: Hiba
authors.initials: H
authors.lastname: Komati
authors.affiliation: From the Laboratory of Molecular Genetics and Cardiac Regeneration, Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Ontario K1N 6N5 and the Graduate Program in Molecular Biology, Institut de Recherches Cliniques de Montréal (IRCM), Université de Montréal, Montréal, Québec H2W 1R7, Canada mona.nemer@uottawa.ca.
authors.forename: Mona
authors.initials: M
authors.lastname: Nemer
mesh_terms: D000015:Abnormalities, Multiple / Q000235:genetics; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005091:Exons; D015870:Gene Expression; D006330:Heart Defects, Congenital / Q000235:genetics; D006344:Heart Septal Defects, Atrial / Q000235:genetics; D006801:Humans; D038061:Lower Extremity Deformities, Congenital / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D032342:Muscle Cells / Q000166:cytology / Q000378:metabolism; D018482:Muscle, Skeletal / Q000254:growth & development / Q000378:metabolism; D009154:Mutation; D009206:Myocardium / Q000378:metabolism / Q000648:ultrastructure; D020033:Protein Isoforms / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D020825:T-Box Domain Proteins / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D038062:Upper Extremity Deformities, Congenital / Q000235:genetics
chemical_list: D020033:Protein Isoforms; D020825:T-Box Domain Proteins; C103172:T-box transcription factor 5
keywords: Alternative Splicing; Gene Regulation; Heart; Muscle Differentiation; Myogenesis; Transcription; Transcription Factors