KIF5B transports BNIP-2 to regulate p38 mitogen-activated protein kinase activation and myoblast differentiation.
The Cdo-p38MAPK (p38 mitogen-activated protein kinase) signaling pathway plays important roles in regulating skeletal myogenesis. During myogenic differentiation, the cell surface receptor Cdo bridges scaffold proteins BNIP-2 and JLP and activates p38MAPK, but the spatial-temporal regulation of this process is largely unknown. We here report that KIF5B, the heavy chain of kinesin-1 motor, is a novel interacting partner of BNIP-2. Coimmunoprecipitation and far-Western study revealed that BNIP-2 directly interacted with the motor and tail domains of KIF5B via its BCH domain. By using a range of organelle markers and live microscopy, we determined the endosomal localization of BNIP-2 and revealed the microtubule-dependent anterograde transport of BNIP-2 in C2C12 cells. The anterograde transport of BNIP-2 was disrupted by a dominant-negative mutant of KIF5B. In addition, knockdown of KIF5B causes aberrant aggregation of BNIP-2, confirming that KIF5B is critical for the anterograde transport of BNIP-2 in cells. Gain- and loss-of-function experiments further showed that KIF5B modulates p38MAPK activity and in turn promotes myogenic differentiation. Of importance, the KIF5B-dependent anterograde transport of BNIP-2 is critical for its promyogenic effects. Our data reveal a novel role of KIF5B in the spatial regulation of Cdo-BNIP-2-p38MAPK signaling and disclose a previously unappreciated linkage between the intracellular transporting system and myogenesis regulation.
Center for Stem Cell and Tissue Engineering, Department of Biochemistry and Molecular Biology, and Zhejiang Provincial Key Lab for Tissue Engineering and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.
Peng
P
Yi
Department of Biological Sciences and Mechanobiology Institute, National University of Singapore, 117411 Singapore.
Li Li
LL
Chew
Center for Stem Cell and Tissue Engineering, Department of Biochemistry and Molecular Biology, and Zhejiang Provincial Key Lab for Tissue Engineering and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.
Ziwang
Z
Zhang
Zhejiang Provincial Key Lab for Tissue Engineering and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.
Hao
H
Ren
Center for Stem Cell and Tissue Engineering, Department of Biochemistry and Molecular Biology, and Zhejiang Provincial Key Lab for Tissue Engineering and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.
Feiya
F
Wang
Center for Stem Cell and Tissue Engineering, Department of Biochemistry and Molecular Biology, and Zhejiang Provincial Key Lab for Tissue Engineering and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.
Xiaoxia
X
Cong
Center for Stem Cell and Tissue Engineering, Department of Biochemistry and Molecular Biology, and.
Liling
L
Zheng
Center for Stem Cell and Tissue Engineering, Department of Biochemistry and Molecular Biology, and.
Yan
Y
Luo
Zhejiang Provincial Key Lab for Tissue Engineering and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.
Hongwei
H
Ouyang
Department of Biological Sciences and Mechanobiology Institute, National University of Singapore, 117411 Singapore zhouyt@zju.edu.cn dbslowbc@nus.edu.sg.
Boon Chuan
BC
Low
Center for Stem Cell and Tissue Engineering, Department of Biochemistry and Molecular Biology, and Zhejiang Provincial Key Lab for Tissue Engineering and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China zhouyt@zju.edu.cn dbslowbc@nus.edu.sg.
Yi Ting
YT
Zhou
D000595:Amino Acid Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D011992:Endosomes / Q000378:metabolism; D006801:Humans; D016547:Kinesins / Q000235:genetics / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D008870:Microtubules / Q000378:metabolism; D008969:Molecular Sequence Data; D024510:Muscle Development; D032446:Myoblasts / Q000166:cytology*; D011485:Protein Binding; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*
C499184:BNIP2 protein, human; D002352:Carrier Proteins; C488101:KIF5B protein, human; D048051:p38 Mitogen-Activated Protein Kinases; D016547:Kinesins