Membrane Protrusion Formation Mediated by Rho/ROCK Signalling and Modulation of Chloride Flux
Membrane Protrusion Formation Mediated by Rho/ROCK Signalling and Modulation of Chloride Flux
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DOI：
10.1101/600379
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发表时间：
2019-04
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影响因子：
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通讯作者：
Akiko Hori;Kenji Nishide;Yuki Yasukuni;K. Haga;Wataru Kakuta;Yasuyuki Ishikawa;M. Hayes;S. Ohnuma;H. Kiyonari;K. Kimura;T. Kondo;Noriaki Sasai
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作者：
Akiko Hori;Kenji Nishide;Yuki Yasukuni;K. Haga;Wataru Kakuta;Yasuyuki Ishikawa;M. Hayes;S. Ohnuma;H. Kiyonari;K. Kimura;T. Kondo;Noriaki Sasai
膜突起是与各种细胞功能相关的重要结构特性。已知pentaspan膜蛋白Prominin-1（Prom 1/CD 133）定位于突起，并在迁移和细胞形态的确定中起关键作用;然而，其潜在机制一直难以捉摸。在这里，我们证明了Prom 1足以触发膜突起的形成。Prom 1在RPE-1细胞中的过表达触发多个长的富含胆固醇的突起，独立于肌动蛋白和微管蛋白聚合。对于这种突起的形成，位于羧基胞质区域的五个氨基酸延伸是必不可少的。此外，小的GT-Rho及其效应激酶ROCK对于这种突起的形成是必不可少的，并且活性Rho和Prom 1的交叉点是突起形成的起始点。重要的是，Prom 1导致由钙离子摄取诱导的氯离子流出，并且突起形成与氯离子流出活性密切相关。总之，这项研究阐明了Prom 1起着关键作用的膜形态和氯离子通量。
Membrane protrusion is an important structural property associated with various cellular functions. The pentaspan membrane protein Prominin-1 (Prom1/CD133) is known to be localised to the protrusions and plays a pivotal role in migration and the determination of cellular morphology; however, the underlying mechanisms have been elusive. Here, we demonstrate that Prom1 is sufficient to trigger membrane protrusion formation. Overexpression of Prom1 in the RPE-1 cells triggers multiple long cholesterol-enriched protrusions, independently from actin and tubulin polymerisation. For this protrusion formation, the five amino acid stretch located at the carboxyl cytosolic region is essential. Moreover, the small GTPase Rho and its effector kinase ROCK are essential for this protrusion formation, and the intersection point of active Rho and Prom1 is where the protrusion formation initiates. Importantly, Prom1 causes the chloride ion efflux induced by calcium ion uptake, and protrusion formation is closely associated with the chloride efflux activity. Altogether, this study has elucidated that Prom1 plays critical roles for the membrane morphology and chloride ion flux.