Akt Regulates a Rab11-Effector Switch Required for Ciliogenesis.

Developmental cell (2019-06-18)
Vijay Walia, Adrian Cuenca, Melanie Vetter, Christine Insinna, Sumeth Perera, Quanlong Lu, Daniel A Ritt, Elizabeth Semler, Suzanne Specht, Jimmy Stauffer, Deborah K Morrison, Esben Lorentzen, Christopher J Westlake
要旨

Serum starvation stimulates cilia growth in cultured cells, yet serum factors associated with ciliogenesis are unknown. Previously, we showed that starvation induces rapid Rab11-dependent vesicular trafficking of Rabin8, a Rab8 guanine-nucleotide exchange factor (GEF), to the mother centriole, leading to Rab8 activation and cilium growth. Here, we demonstrate that through the LPA receptor 1 (LPAR1), serum lysophosphatidic acid (LPA) inhibits Rab11a-Rabin8 interaction and ciliogenesis. LPA/LPAR1 regulates ciliogenesis initiation via downstream PI3K/Akt activation, independent of effects on cell cycle. Akt stabilizes Rab11a binding to its effector, WDR44, and a WDR44-pAkt-phosphomimetic mutant blocks ciliogenesis. WDR44 depletion promotes Rabin8 preciliary trafficking and ciliogenesis-initiating events at the mother centriole. Our work suggests disruption of Akt signaling causes a switch from Rab11-WDR44 to the ciliogenic Rab11-FIP3-Rabin8 complex. Finally, we demonstrate that Akt regulates downstream ciliogenesis processes associated with Rab8-dependent cilia growth. Together, this study uncovers a mechanism whereby serum mitogen signaling regulates Rabin8 preciliary trafficking and ciliogenesis initiation.

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製品内容

Sigma-Aldrich
hEGF, EGF, recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture
Sigma-Aldrich
モノクロナール抗アセチル化チューブリン マウス宿主抗体, clone 6-11B-1, purified from hybridoma cell culture
Sigma-Aldrich
ドキシサイクリン塩酸塩、調製済み溶液
Sigma-Aldrich
モノクロナール抗チューブリン, アセチル化 マウス宿主抗体, clone 6-11B-1, ascites fluid
Sigma-Aldrich
モノクロナール抗β-アクチン−ペルオキシダーゼ マウス宿主抗体, clone AC-15, purified from hybridoma cell culture
Roche
抗HAペルオキシダーゼ、High Affinity, from rat IgG1
Sigma-Aldrich
モノクロナール抗γ-チューブリン マウス宿主抗体, clone GTU-88, ascites fluid
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ワートマニン, from Penicillium funiculosum, ≥98% (HPLC and TLC)
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3-sn-ホスファチジン酸 ナトリウム塩 from egg yolk lecithin, ≥98%
Sigma-Aldrich
GSK690693, ≥98% (HPLC)
Sigma-Aldrich
Monoclonal Anti-LPAR1 antibody produced in mouse, clone 2E2, purified immunoglobulin, buffered aqueous solution