A complex of Rab13 with MICAL-L2 and α-actinin-4 is essential for insulin-dependent GLUT4 exocytosis
In: Molecular Biology of the Cell, Jg. 27 (2016), S. 75-89
Online
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Zugriff:
Rab13 is necessary for insulin-regulated GLUT4-vesicle exocytosis in muscle. Biochemical and imaging analyses provide evidence that activated Rab13 engages a scaffold protein MICAL-L2 to form a complex with Rab13 and α-actinin-4. Through GLUT4 interaction with α-actinin-4, GLUT4 vesicles are recruited to the muscle plasma membrane.
Insulin promotes glucose uptake into skeletal muscle through recruitment of glucose transporter 4 (GLUT4) to the plasma membrane. Rab GTPases are molecular switches mobilizing intracellular vesicles, and Rab13 is necessary for insulin-regulated GLUT4–vesicle exocytic translocation in muscle cells. We show that Rab13 engages the scaffold protein MICAL-L2 in this process. RNA interference–mediated knockdown of MICAL-L2 or truncated MICAL-L2 (MICAL-L2-CT) impaired insulin-stimulated GLUT4 translocation. Insulin increased Rab13 binding to MICAL-L2, assessed by pull down and colocalization under confocal fluorescence and structured illumination microscopies. Association was also visualized at the cell periphery using TIRF microscopy. Insulin further increased binding of MICAL-L2 to α-actinin-4 (ACTN4), a protein involved in GLUT4 translocation. Rab13, MICAL-L2, and ACTN4 formed an insulin-dependent complex assessed by pull down and confocal fluorescence imaging. Of note, GLUT4 associated with the complex in response to insulin, requiring the ACTN4-binding domain in MICAL-L2. This was demonstrated by pull down with distinct fragments of MICAL-L2 and confocal and structured illumination microscopies. Finally, expression of MICAL-L2-CT abrogated the insulin-dependent colocalization of Rab13 with ACTN4 or Rab13 with GLUT4. Our findings suggest that MICAL-L2 is an effector of insulin-activated Rab13, which links to GLUT4 through ACTN4, localizing GLUT4 vesicles at the muscle cell periphery to enable their fusion with the membrane.
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A complex of Rab13 with MICAL-L2 and α-actinin-4 is essential for insulin-dependent GLUT4 exocytosis
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Autor/in / Beteiligte Person: | Bilan, Philip J. ; Klip, Amira ; Jaldin-Fincati, Javier R. ; Liu, Zhi ; Sun, Yi |
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Zeitschrift: | Molecular Biology of the Cell, Jg. 27 (2016), S. 75-89 |
Veröffentlichung: | American Society for Cell Biology (ASCB), 2016 |
Medientyp: | unknown |
ISSN: | 1939-4586 (print) ; 1059-1524 (print) |
DOI: | 10.1091/mbc.e15-05-0319 |
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