Par1b/MARK2 phosphorylates kinesin-like motor protein GAKIN/KIF13B to regulate axon formation.
Here we report that Par1b/MARK2 regulates axon formation via phosphorylation of a kinesin superfamily protein GAKIN/KIF13B. Accumulating evidence indicated the importance of the evolutionarily conserved kinase Par1b in the regulation of cell polarity. Using hippocampal neurons in culture, it has been shown that Par1b regulates axon specification, but the underlying mechanism remains uncharacterized. We identify GAKIN/KIF13B as a novel Par1b-binding protein and reveal that GAKIN/KIF13B is a physiological substrate for Par1b, and the phosphorylation sites are conserved from Drosophila. In hippocampal neurons, GAKIN/KIF13B accumulates at the distal part of the microtubules in the tips of axons, but not of dendrites. Overexpression of GAKIN/KIF13B by itself can induce the formation of extra axons, which is inhibited by the coexpression of Par1b in a manner dependent on its kinase activity. In contrast, small interfering RNA (siRNA)-mediated knockdown of GAKIN/KIF13B severely retards neurite extension and promotes the axonless phenotype. The extra axon phenotype caused by Par1b siRNA is suppressed by cointroduction of GAKIN/KIF13B siRNA, thus placing the GAKIN/KIF13B function downstream of Par1b. We also find that GAKIN/KIF13B acts downstream of the phosphatidylinositol 3-kinase (PI3K) signaling via Par1b phosphorylation. These results reveal that GAKIN/KIF13B is a key intermediate linking Par1b to the regulation of axon formation.
Yoshimura
Yuta
Y
Laboratory of Intracellular Signaling, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Terabayashi
Takeshi
T
Miki
Hiroaki
H
D048948:14-3-3 Proteins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001369:Axons / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D016547:Kinesins / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D020409:Molecular Motor Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D021381:Protein Transport; D051381:Rats
D048948:14-3-3 Proteins; D016547:Kinesins; D020409:Molecular Motor Proteins; D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; C094670:MARK2 protein, human; C105549:MARK2 protein, rat; C513406:KIF13B protein, human; C504977:KIF13B protein, rat