Phosphorylation on threonine 11 of [beta]-dystrobrevin alters its interaction with kinesin heavy chain

• Published in: The FEBS journal Vol. 279; no. 22; p. 4131
• Main Authors: Fratini, Federica, Macchia, Gianfranco, Torreri, Paola, Matteucci, Andrea, Salzano, Anna Maria, Crescenzi, Marco, Macioce, Pompeo, Petrucci, Tamara C, Ceccarini, Marina
• Format: Journal Article
• Language: English
• Published: Oxford Blackwell Publishing Ltd 01.11.2012
• Subjects: Cellular biology; Kinases; Molecular biology; Proteins
• ISSN: 1742-464X; 1742-4658
• DOI: 10.1111/febs.12006

Dystrobrevin family members ([alpha] and [beta]) are cytoplasmic components of the dystrophin-associated glycoprotein complex, a multimeric protein complex first isolated from skeletal muscle, which links the extracellular matrix to the actin cytoskeleton. Dystrobrevin shares high homology with the cysteine-rich and C-terminal domains of dystrophin and a common domain organization. The [beta]-dystrobrevin isoform is restricted to nonmuscle tissues, serves as a scaffold for signaling complexes, and may participate in intracellular transport through its interaction with kinesin heavy chain. We have previously characterized the molecular determinants affecting the [beta]-dystrobrevin-kinesin heavy chain interaction, among which is cAMP-dependent protein kinase [protein kinase A (PKA)] phosphorylation of [beta]-dystrobrevin. In this study, we have identified [beta]-dystrobrevin residues phosphorylated in vitro by PKA with pull-down assays, surface plasmon resonance measurements, and MS analysis. Among the identified phosphorylated residues, we demonstrated, by site-directed mutagenesis, that Thr11 is the regulatory site for the [beta]-dystrobrevin-kinesin interaction. As dystrobrevin may function as a signaling scaffold for kinases/phosphatases, we also investigated whether [beta]-dystrobrevin is phosphorylated in vitro by kinases other than PKA. Thr11 was phosphorylated by protein kinase C, suggesting that this represents a key residue modified by the activation of different signaling pathways. Structured digital abstract [PUBLICATION ABSTRACT]