The LIM-only protein FHL2 interacts with beta-catenin and promotes differentiation of mouse myoblasts

The Journal of Cell Biology
Bernd MartinViktor Wixler

Abstract

FHL2 is a LIM-domain protein expressed in myoblasts but down-regulated in malignant rhabdomyosarcoma cells, suggesting an important role of FHL2 in muscle development. To investigate the importance of FHL2 during myoblast differentiation, we performed a yeast two-hybrid screen using a cDNA library derived from myoblasts induced for differentiation. We identified beta-catenin as a novel interaction partner of FHL2 and confirmed the specificity of association by direct in vitro binding tests and coimmunoprecipitation assays from cell lysates. Deletion analysis of both proteins revealed that the NH2-terminal part of beta-catenin is sufficient for binding in yeast, but addition of the first armadillo repeat is necessary for binding FHL2 in mammalian cells, whereas the presence of all four LIM domains of FHL2 is needed for the interaction. Expression of FHL2 counteracts beta-catenin-mediated activation of a TCF/LEF-dependent reporter gene in a dose-dependent and muscle cell-specific manner. After injection into Xenopus embryos, FHL2 inhibited the beta-catenin-induced axis duplication. C2C12 mouse myoblasts stably expressing FHL2 show increased myogenic differentiation reflected by accelerated myotube formation and expression of musc...Continue Reading

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Methods Mentioned

BETA
two-hybrid
fluorescence microscopy
transfection
transfections
pull-down
coimmunoprecipitation
PCR
FCS

Related Concepts

Fhl2 protein, mouse
Beta-catenin protein, Xenopus
Catnb protein, mouse
Metazoa
Cell Differentiation Process
Cytoskeletal Proteins
Immunocytochemistry
Muscle Proteins
Oocytes
Hybrid Proteins, Recombinant

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