Sox6 regulation of cardiac myocyte development

• Published in: Nucleic acids research 2003-10, Vol.31 (20), p.5941-5948
• Main Authors: Cohen‐Barak, Orit, Yi, Zanhua, Hagiwara, Nobuko, Monzen, Koshiro, Komuro, Issei, Brilliant, Murray H.
• Format: Article
• Language: English
• Subjects: Animals; Blotting, Northern; Calcium Channels, L-Type - genetics; Calcium Channels, L-Type - metabolism; Cell Differentiation - genetics; Cell Differentiation - physiology; Cell Line, Tumor; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Female; Gene Expression; Genotype; High Mobility Group Proteins - genetics; High Mobility Group Proteins - metabolism; Humans; Male; Mice; Mice, Mutant Strains; Mutation; Myocytes, Cardiac - cytology; Myocytes, Cardiac - metabolism; noggin; Peptides - genetics; Peptides - metabolism; Proline-Rich Protein Domains; Protein Binding; Prtb protein; RNA, Messenger - genetics; RNA, Messenger - metabolism; SOXD Transcription Factors; Transcription Factors; Two-Hybrid System Techniques
• ISSN: 0305-1048; 1362-4962; 1362-4962
• DOI: 10.1093/nar/gkg807

A mouse mutation (p100H/p100H) has been identified that is associated with cardioskeletal myopathy, heart block, delayed growth and early postnatal death. The gene that is disrupted in this mutation encodes the transcription factor Sox6. P19CL6 cells were used as an in vitro cardiomyocyte differentiation system and revealed that Sox6 is expressed exclusively when the cells are committed to differentiate to beating cardiac myocytes. We used the yeast two‐hybrid system to identify the Prtb (Proline‐rich transcript of the brain) protein as a Sox6 interactor, and subsequently confirmed the interaction by co‐immunoprecipitation. Prtb expression in P19CL6 cells increased with differentiation to beating cardiomyocytes. Using the P19CL6 cells stably transfected with noggin, an antagonist of BMP (Bone Morphogenic Protein), we found that BMP expression is required for Sox6 expression in cardiomyocyte differentiation. Surprisingly, the expression of the α1c‐subunit gene of the L‐type Ca2+ channel decreased in P19CL6 cells as they differentiated to beating cardiac cells. Ectopic expression of Sox6 or Prtb alone in P19CL6 cells caused down‐regulation of L‐type Ca2+ α1c expression, but when Sox6 and Prtb were co‐transfected to the cells, L‐type Ca2+ α1c remained at basal levels. A similar relationship of Sox6 and L‐type Ca2+ α1c expression was seen in vivo (comparing wild‐type and p100H/p100H mutant mice). Thus, Sox6 is within the BMP pathway in cardiac differentiation, interacts with Prtb and may play a critical role in the regulation of a cardiac L‐type Ca2+ channel.