Dynamic analysis of Arabidopsis AP2 sigma subunit reveals a key role in clathrin-mediated endocytosis and plant development

doi:10.1242/dev.095711

	Dynamic analysis of Arabidopsis AP2 sigma subunit reveals a key role in clathrin-mediated endocytosis and plant development
	Fan, Lusheng 1; Hao, Huaiqing ; Xue, Yiqun 1; Zhang, Liang 2; Song, Kai 1; Ding, Zhaojun 3; Botella, Miguel A.; Wang, Haiyang 5; Lin, Jinxing
	2013
发表期刊	DEVELOPMENT
ISSN	0950-1991
卷号	140 期号:18 页码:3826-3837
摘要	Clathrin-mediated endocytosis, which depends on the AP2 complex, plays an essential role in many cellular and developmental processes in mammalian cells. However, the function of the AP2 complex in plants remains largely unexplored. Here, we show in Arabidopsis that the AP2 sigma subunit mutant (ap2 sigma) displays various developmental defects that are similar to those of mutants defective in auxin transport and/or signaling, including single, trumpet-shaped and triple cotyledons, impaired vascular pattern, reduced vegetative growth, defective silique development and drastically reduced fertility. We demonstrate that AP2 sigma is closely associated and physically interacts with the clathrin light chain (CLC) in vivo using fluorescence cross-correlation spectroscopy (FCCS), protein proximity analyses and co-immunoprecipitation assays. Using variable-angle total internal reflection fluorescence microscopy (VA-TIRFM), we show that AP2 sigma-mCherry spots colocalize with CLC-EGFP at the plasma membrane, and that AP2 s-mCherry fluorescence appears and disappears before CLC-EGFP fluorescence. The density and turnover rate of the CLC-EGFP spots are significantly reduced in the ap2 sigma mutant. The internalization and recycling of the endocytic tracer FM4-64 and the auxin efflux carrier protein PIN1 are also significantly reduced in the ap2 sigma mutant. Further, the polar localization of PIN1-GFP is significantly disrupted during embryogenesis in the ap2 sigma mutant. Taken together, our results support an essential role of AP2 s in the assembly of a functional AP2 complex in plants, which is required for clathrin-mediated endocytosis, polar auxin transport and plant growth regulation.
关键词	AP2 complex AP-2 Clathrin-mediated endocytosis FCCS PIN auxin transporter VA-TIRFM
学科领域	Developmental Biology
DOI	10.1242/dev.095711
收录类别	SCI
语种	英语
WOS关键词	REFLECTION FLUORESCENCE MICROSCOPY ; CORRELATION SPECTROSCOPY ; ADAPTER COMPLEX ; RECEPTOR ; BINDING ; PROTEINS ; AP-2 ; NETWORK ; GROWTH ; DOMAIN
WOS研究方向	Science Citation Index Expanded (SCI-EXPANDED)
WOS记录号	WOS:000323698100012
出版者	COMPANY BIOLOGISTS LTD
文献子类	Article
出版地	CAMBRIDGE
EISSN	1477-9129
资助机构	National Basic Research Program of China(National Basic Research Program of China) ; project Program for Creative Research Teams ; National Natural Science Foundation of China(National Natural Science Foundation of China (NSFC))
作者邮箱	Botella, Miguel Ángel/S-3676-2019 ; Botella, Miguel A/B-5734-2012
引用统计	被引频次：115[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.ibcas.ac.cn/handle/2S10CLM1/28019
专题	中科院植物分子生理学重点实验室
作者单位	1.Chinese Acad Sci, Inst Bot, Key Lab Plant Mol Physiol, Beijing 100093, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China 3.Henan Normal Univ, Coll Life Sci, Xinxiang 453007, Peoples R China 4.Shandong Univ, Sch Life Sci, Jinan 250100, Peoples R China 5.Botella, Miguel A.] Univ Malaga, Dept Biol Celular Genet & Fisiol, E-29071 Malaga, Spain 6.Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA
推荐引用方式 GB/T 7714	Fan, Lusheng,Hao, Huaiqing,Xue, Yiqun,et al. Dynamic analysis of Arabidopsis AP2 sigma subunit reveals a key role in clathrin-mediated endocytosis and plant development[J]. DEVELOPMENT,2013,140(18):3826-3837.
APA	Fan, Lusheng.,Hao, Huaiqing.,Xue, Yiqun.,Zhang, Liang.,Song, Kai.,...&Lin, Jinxing.(2013).Dynamic analysis of Arabidopsis AP2 sigma subunit reveals a key role in clathrin-mediated endocytosis and plant development.DEVELOPMENT,140(18),3826-3837.
MLA	Fan, Lusheng,et al."Dynamic analysis of Arabidopsis AP2 sigma subunit reveals a key role in clathrin-mediated endocytosis and plant development".DEVELOPMENT 140.18(2013):3826-3837.

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