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Ankrd1 inhibits the FAK/Rho-GTPase/F-actin pathway by downregulating ITGA6 transcriptional to regulate myoblast functions

J Cell Physiol. 2024 Nov;239(11):e31359. doi: 10.1002/jcp.31359. Epub 2024 Jul 10.

Abstract

Skeletal muscle constitutes the largest percentage of tissue in the animal body and plays a pivotal role in the development of normal life activities in the organism. However, the regulation mechanism of skeletal muscle growth and development remains largely unclear. This study investigated the effects of Ankrd1 on the proliferation and differentiation of C2C12 myoblasts. Here, we identified Ankrd1 as a potential regulator of muscle cell development, and found that Ankrd1 knockdown resulted in the proliferation ability decrease but the differentiation level increase of C2C12 cells. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyzes as well as RNA-seq results showed that Ankrd1 knockdown activated focal adhesion kinase (FAK)/F-actin signal pathway with most genes significantly enriched in this pathway upregulated. The integrin subunit Itga6 promoter activity is increased when Ankrd1 knockdown, as demonstrated by a dual-luciferase reporter assay. This study revealed the molecular mechanism by which Ankrd1 knockdown enhanced FAK phosphorylation activity through the alteration of integrin subunit levels, thus activating FAK/Rho-GTPase/F-actin signal pathway, eventually promoting myoblast differentiation. Our data suggested that Ankrd1 might serve as a potential regulator of muscle cell development. Our findings provide new insights into skeletal muscle growth and development and valuable references for further study of human muscle-related diseases.

Keywords: Ankrd1; C2C12 myoblast; FAK; F‐actin; differentiation; integrin.

MeSH terms

  • Actins* / genetics
  • Actins* / metabolism
  • Animals
  • Cell Differentiation* / genetics
  • Cell Line
  • Cell Proliferation* / genetics
  • Down-Regulation / genetics
  • Focal Adhesion Kinase 1* / genetics
  • Focal Adhesion Kinase 1* / metabolism
  • Humans
  • Integrin alpha6* / genetics
  • Integrin alpha6* / metabolism
  • Mice
  • Muscle Development / genetics
  • Muscle Proteins
  • Muscle, Skeletal / metabolism
  • Myoblasts* / metabolism
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Phosphorylation
  • Repressor Proteins* / genetics
  • Repressor Proteins* / metabolism
  • Signal Transduction*
  • rho GTP-Binding Proteins / genetics
  • rho GTP-Binding Proteins / metabolism

Substances

  • Repressor Proteins
  • Actins
  • Focal Adhesion Kinase 1
  • Integrin alpha6
  • Nuclear Proteins
  • ANKRD1 protein, human
  • ITGA6 protein, human
  • rho GTP-Binding Proteins
  • PTK2 protein, human
  • Muscle Proteins