Post-Golgi trafficking of the mannose 6-phosphate receptor in living cells

活细胞中甘露糖 6-磷酸受体的后高尔基体运输

• 批准号: 17590163
• 负责人: WAGURI Satoshi
• 金额: $ 2.24万
• 依托单位: Fukushima Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590163/
• 关键词: mannose 6-phosphate receptors; clathrin adaptors; GGAs; I-cell disease; trans-Golgi network; lysosomal enzyme; AP1; ライブセルイメージング; フォトブリーチ

1. Sorting of lysosomal enzymes is conducted by mannose 6-phosphate receptors (M6PR), which are transported between the trans-Golgi network (TGN), endosomes, and plasma membrane. In this research project, we examined functions of the extracytoplasmic domain of M6PR in its intracellular trafficking, and obtained the following conclusions.(1) Time-lapse imaging and FRAP (fluorescence recovery after photobleaching) analyses of GFP-M6PR, together with a chloroquine treatment suggest that the extracytoplasmic domain of M6PR is required for tight interaction with endocytic compartments and retention by them.(2) Experiments using a drug, U18666A, and fibroblasts derived from I-cell disease patients, suggest that the binding to the M6P ligands is involved in yet unidentified transport steps of M6PR. U18666A could be a useful tool to investigate new transport steps of M6PR.2. To investigate the function of clathrin adaptors, GGAs, that regulate the M6PR trafficking, we performed RNAi and dominant-negative experiments for GGA2. Although the depletion of GGA2 alone did not cause an apparent re distribution of M6PR, a lysosomal enzyme, cathepsin D, was significantly missecreted. These results suggest that missorting of lysosomal enzymes is not always accompanied by the drastic M6PR redistribution. We will perform detailed analyses as to which transport steps are regulated by GGAs.

1.溶酶体酶的分选通过甘露糖6-磷酸受体（M6 PR）进行，其在反式高尔基体网络（TGN）、内体和质膜之间转运。在本研究项目中，我们研究了M6 PR胞质外结构域在其胞内运输中的功能，并获得了以下结论。(1)GFP-M6 PR的延时成像和FRAP（光漂白后的荧光恢复）分析以及氯喹处理表明，M6 PR的胞质外结构域是与胞吞隔室紧密相互作用并被它们保留所必需的。(2)使用药物U18666 A和来自I细胞疾病患者的成纤维细胞的实验表明，与M6 P配体的结合涉及M6 PR的尚未鉴定的转运步骤。U18666 A可能是研究M6PR.2新的转运步骤的有用工具。为了研究调节M6 PR运输的网格蛋白衔接子GGA的功能，我们对GGA 2进行了RNAi和显性阴性实验。尽管GGA 2单独消耗不会引起M6 PR的明显重新分布，但溶酶体酶组织蛋白酶D显著错误分泌。这些结果表明，溶酶体酶的分选错误并不总是伴随着剧烈的M6 PR再分布。我们将详细分析GGA对哪些运输步骤进行监管。

项目成果

Loss of autophagy in the central nervous system causes neurodegeneration in mice

• DOI: 10.1038/nature04723
• 发表时间: 2006-06-15
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Komatsu, Masaaki;Waguri, Satoshi;Tanaka, Keiji
• 通讯作者: Tanaka, Keiji

Uncoupling protein 2 negatively regulates neurite extensions in PC12h cells.

解偶联蛋白 2 负向调节 PC12h 细胞中的神经突延伸。

• 发表时间: 2006
• 期刊: Neurosci Lett 410(2)
• 作者: Yamada S;Isojima Y;Kanamori S;Waguri S;Uchiyama Y;Nagai K.
• 通讯作者: Nagai K.

Participation of autophagy in storage of lysosomes in neurons from mouse models of neuronal ceroid-lipofuscinoses (Batten disease)

• DOI: 10.1016/s0002-9440(10)61253-9
• 发表时间: 2005-12-01
• 期刊: AMERICAN JOURNAL OF PATHOLOGY
• 影响因子: 6
• 作者: Koike, M;Shibata, M;Uchiyama, Y
• 通讯作者: Uchiyama, Y

C‐terminal Src kinase controls development and maintenance of mouse squamous epithelia

• DOI: 10.1038/sj.emboj.7601595
• 发表时间: 2007-03
• 期刊: The EMBO Journal
• 作者: R. Yagi;S. Waguri;Y. Sumikawa;S. Nada;Chitose Oneyama;S. Itami;C. Schmedt;Y. Uchiyama;M. Okada

Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice.

• DOI: 10.1083/jcb.200412022
• 发表时间: 2005-05-09
• 期刊: The Journal of cell biology
• 作者: Komatsu M;Waguri S;Ueno T;Iwata J;Murata S;Tanida I;Ezaki J;Mizushima N;Ohsumi Y;Uchiyama Y;Kominami E;Tanaka K;Chiba T
• 通讯作者: Chiba T