Cracknell, Tobias, Mannsverk, Steinar, Nichols, Angus et al. (2 more authors) (2020) Proteomic resolution of IGFN1 complexes reveals a functional interaction with the actin nucleating protein COBL. Experimental cell research. 112179. ISSN 0014-4827
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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | /© 2020 University of York, UK. |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Biology (York) The University of York > Faculty of Sciences (York) > Chemistry (York) |
Depositing User: | Pure (York) |
Date Deposited: | 21 Jul 2020 16:10 |
Last Modified: | 16 Oct 2024 16:47 |
Published Version: | https://doi.org/10.1016/j.yexcr.2020.112179 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1016/j.yexcr.2020.112179 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:163470 |
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Description: Proteomic resolution of IGFN1 complexes reveals a functional interaction with the actin nucleating protein COBL
Licence: CC-BY-NC-ND 2.5