Muscle LIM protein/CSRP3: a mechanosensor with a role in autophagy

Cell Death Discovery
M M RashidMarco Tafani

Abstract

Muscle LIM protein (MLP) is a microtubule-associated protein expressed in cardiac and muscle tissues that belongs to the cysteine-rich protein (CSRP/CRP) family. MLP has a central role during muscle development and for architectural maintenance of muscle cells. However, muscle cells rely on autophagy during differentiation and for structural maintenance. To study the role of MLP in autophagy, we have used C2C12 mouse myoblasts silenced or overexpressing MLP. Our results show that MLP contributes to the correct autophagosome formation and flux by interacting with LC3 as demonstrated by co-immunoprecipitation and PLA assay. In fact, MLP silencing results in decreased LC3-II staining and absent degradation of long-lived proteins. Moreover, MLP silencing impaired myoblasts differentiation as measured by decreased expression of MyoD1, MyoG1 and myosin heavy chain. Ultrastructural analysis revealed the presence of large empty autophagosomes in myoblasts and multimembranous structures in myotubes from MLP-silenced clones. Impaired autophagy in MLP-silenced cells resulted in increased susceptibility to apoptotic cell death. In fact, treatment of MLP-silenced C2C12 myoblasts and myotubes with staurosporine resulted in increased caspase-...Continue Reading

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Citations

Jan 26, 2020·International Journal of Molecular Sciences·Can CuiAnd Huadong Yin
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Methods Mentioned

BETA
co-immunoprecipitation
immunoprecipitation
proximity ligation assay
transmission electron microscopy
flow cytometry
lipidation
co-immunoprecipitation assay
light microscopy
transfecting
PCR

Related Concepts

Analysis of Substances
Autophagy
C-reactive protein
Cell Death
Cell Differentiation Process
Disease Susceptibility
Heart
Microtubule-Associated Proteins
Muscle
Myosin Heavy Chains

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