Spatiotemporal Localization of Rab20 in Live RAW264 Macrophages during Macropinocytosis.

Spatiotemporal Localization of Rab20 in Live RAW264 Macrophages during Macropinocytosis.
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DOI:
10.1267/ahc.12014
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发表时间:
2012-12-26
影响因子:
2.4
通讯作者:
Araki N
Araki N
中科院分区:
生物学4区
文献类型:
--
作者:
Egami Y;Araki N

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Rab 20是Rab GT3家族的成员,但其在巨胞饮中的意义尚不清楚。我们通过荧光蛋白融合Rab 20的活细胞成像研究Rab 20在RAW 264巨噬细胞中的时空定位。结果表明,Rab 20与大胞饮体膜短暂结合。在巨胞饮体形成的早期阶段,Rab 20略微定位于圆形皱褶(巨胞饮杯),巨胞饮体的前体形式,并越来越多地被招募到新形成的巨胞饮体。虽然Rab 20与Rab 5和Rab 21共定位在巨胞饮体膜上,但Rab 20与巨胞饮体的结合即使在Rab 5和Rab 21从巨胞饮体膜上解离后也持续存在。然后Rab 20与Rab 7和Lamp 1(晚期内体/溶酶体标记物)在巨胞饮体上共定位一段时间。我们的数据表明,Rab 20是巨胞饮途径的一个新的组成部分,并在从早期到晚期巨胞饮体成熟的长期阶段发挥作用。
Rab20 is a member of the Rab GTPase family, but its implication in macropinocytosis is unclear. We examined the spatiotemporal localization of Rab20 in RAW264 macrophages by the live-cell imaging of fluorescent protein-fused Rab20. It was shown that Rab20 was transiently associated with macropinosomal membranes. During the early stage of macropinosome formation, Rab20 was slightly localized on the circular ruffles (macropinocytic cups), the precursor forms of macropinosomes, and was increasingly recruited to the newly formed macropinosomes. Although Rab20 was colocalized with Rab5 and Rab21 on macropinosomal membranes, the association of Rab20 with macropinosomes persisted even after the dissociations of Rab5 and Rab21 from macropinosomal membranes. Rab20 was then colocalized with Rab7 and Lamp1, late endosomal/lysosomal markers, on macropinosomes for a while. Our data indicate that Rab20 is a novel component of macropinocytic pathway and functions at long-standing stages from early to late macropinosome maturation.
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