Molecular cloning of TPM3 gene in qinchuan cattle and its effect on myoblast proliferation and differentiation

• Published in: Animal biotechnology 2024-11, Vol.35 (1), p.2345238-2345238
• Main Authors: Guo, Juntao, Wang, Jianfang, Zhang, Ke, Yang, Zhimei, Li, Bingzhi, Pan, Yueting, Yu, Hengwei, Yu, Shengchen, Abbas Raza, Sayed Haidar, Kuraz Abebea, Belete, Zan, Linsen
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; Animals; Bioinformatics; Cattle; Cattle - genetics; Cell differentiation; Cell Differentiation - genetics; Cell Proliferation; Cloning; Cloning, Molecular; Differentiation; Exploratory behavior; Livestock; Livestock breeding; mRNA; Muscle contraction; Muscle Development - genetics; Muscle, Skeletal; Muscles; Muscular function; Musculoskeletal system; Myoblasts; Myoblasts - cytology; Myoblasts - metabolism; Myocardium; Myotubes; Phosphorylation; Polymerase chain reaction; proliferation and differentiation; Protein structure; Proteins; qinchuan cattle; R&D; Real time; Regulatory sequences; Research & development; Secondary structure; Skeletal muscle; TPM3 gene; Tropomyosin; Tropomyosin - chemistry; Tropomyosin - genetics; Tropomyosin - metabolism
• ISSN: 1049-5398; 1532-2378
• DOI: 10.1080/10495398.2024.2345238

Tropomyosin 3 (TPM3) plays a significant role as a regulatory protein in muscle contraction, affecting the growth and development of skeletal muscles. Despite its importance, limited research has been conducted to investigate the influence of TPM3 on bovine skeletal muscle development. Therefore, this study revealed the role of TPM3 in bovine myoblast growth and development. This research involved conducting a thorough examination of the Qinchuan cattle TPM3 gene using bioinformatics tools to examine its sequence and structural characteristics. Furthermore, TPM3 expression was evaluated in various bovine tissues and cells using quantitative real-time polymerase chain reaction (qRT-PCR). The results showed that the coding region of TPM3 spans 855 bp, with the 161st base being the T base, encoding a protein with 284 amino acids and 19 phosphorylation sites. This protein demonstrated high conservation across species while displaying a predominant α-helix secondary structure despite being an unstable acidic protein. Notably, a noticeable increase in TPM3 expression was observed in the longissimus dorsi muscle and myocardium of calves and adult cattle. Expression patterns varied during different stages of myoblast differentiation. Functional studies that involved interference with TPM3 in Qinchuan cattle myoblasts revealed a very significantly decrease in S-phase cell numbers and EdU-positive staining (P