ERK2-mediated phosphorylation of Par3 regulates neuronal polarization.
Axon formation is one of the most important events in neuronal polarization and is regulated by signaling molecules involved in cytoskeletal rearrangement and protein transport. We previously found that Partition-defective 3 (Par3) is associated with KIF3A (kinesin-2) and is transported into the nascent axon in a KIF3A-dependent fashion. Par3 interacts with the Rac-specific guanine nucleotide-exchange factors (GEFs) Tiam1/2, which activate Rac1, and participates in axon formation in cultured hippocampal neurons. However, the regulatory mechanism of the Par3-KIF3A interaction is poorly understood, and the role of Par3 in neuronal polarization in vivo remains elusive. Here, we found that extracellular signal-regulated kinase 2 (ERK2) directly interacts with Par3, that ERK2 phosphorylates Par3 at Ser-1116, and that the phosphorylated Par3 accumulates at the axonal tips in a manner dependent upon ERK2 activity. The phosphorylation of Par3 by ERK2 inhibited the interaction of Par3 with KIF3A but not with the other Par3 partners, including Par6 and aPKC. The phosphomimic mutant of Par3 (Par3-S1116D) showed less binding activity with the KIF3s and slower transport in the axons. The knockdown of Par3 by RNA interference impaired neuronal polarization, which was rescued with RNAi-resistant Par3, but not with the phosphomimic Par3 mutant, in cultured rat hippocampal neurons and mouse cortical projection neurons in vivo. These results suggest that ERK2 phosphorylates Par3 and inhibits its binding with KIF3A, thereby controlling Par3 transport and neuronal polarity.
Department of Cell Pharmacology, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan.
Yasuhiro
Y
Funahashi
Takashi
T
Namba
Shin
S
Fujisue
Norimichi
N
Itoh
Shinichi
S
Nakamuta
Katsuhiro
K
Kato
Akiko
A
Shimada
Chundi
C
Xu
Wei
W
Shan
Tomoki
T
Nishioka
Kozo
K
Kaibuchi
D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000378:metabolism*; D016764:Cell Polarity / Q000502:physiology*; D002522:Chlorocebus aethiops; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D016547:Kinesins / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D009419:Nerve Tissue Proteins; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation; D021381:Protein Transport / Q000502:physiology; D034741:RNA, Small Interfering; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection; D014981:Xenopus
D002352:Carrier Proteins; C087415:KIF3A protein, human; D009419:Nerve Tissue Proteins; C472451:Pard3 protein, rat; D034741:RNA, Small Interfering; D019950:Mitogen-Activated Protein Kinase 1; D016547:Kinesins