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Proteomic resolution of IGFN1 complexes reveals a functional interaction with the actin nucleating protein COBL
- Source :
- Experimental Cell Research
- Publication Year :
- 2020
- Publisher :
- Uppsala universitet, Institutionen för medicinsk biokemi och mikrobiologi, 2020.
-
Abstract
- The Igfn1 gene produces multiple proteins by alternative splicing predominantly expressed in skeletal muscle. Igfn1 deficient clones derived from C2C12 myoblasts show reduced fusion index and morphological differences compared to control myotubes. Here, we first show that G:F actin ratios are significantly higher in differentiating IGFN1-deficient C2C12 myoblasts, suggesting that fusion and differentiation defects are underpinned by deficient actin remodelling. We obtained pull-downs from skeletal muscle with IGFN1 fragments and applied a proteomics approach. The proteomic composition of IGFN1 complexes identified the cytoskeleton and an association with the proteasome as the main networks. The actin nucleating protein COBL was selected for further validation. COBL is expressed in C2C12 myoblasts from the first stages of myoblast fusion but not in proliferating cells. COBL is also expressed in adult muscle and, as IGFN1, localizes to the Z-disc. We show that IGFN1 interacts, stabilizes and colocalizes with COBL and prevents the ability of COBL to form actin ruffles in COS7 cells. COBL loss of function C2C12-derived clones are able to fuse, therefore indicating that COBL or the IGFN1/COBL interaction are not essential for myoblast fusion.
- Subjects :
- 0301 basic medicine
Proteomics
Sarcomeres
Cytoskeleton remodelling
Myoblast
Cellbiologi
Cell- och molekylärbiologi
Muscle Fibers, Skeletal
Cell Communication
Biology
Article
Cell Line
Myoblasts
03 medical and health sciences
Myoblast fusion
0302 clinical medicine
medicine
Myocyte
Animals
Cytoskeleton
Muscle, Skeletal
Actin
Myogenesis
Z-disc
Alternative splicing
Microfilament Proteins
Skeletal muscle
Cell Differentiation
Cell Biology
musculoskeletal system
Cell biology
Actin Cytoskeleton
030104 developmental biology
medicine.anatomical_structure
030220 oncology & carcinogenesis
C2C12
tissues
Cell and Molecular Biology
Subjects
Details
- Language :
- English
- ISSN :
- 00144827
- Database :
- OpenAIRE
- Journal :
- Experimental Cell Research
- Accession number :
- edsair.doi.dedup.....9d4c56b6e2a314141fd8a14fcec02b3c