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

基本信息

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

项目摘要

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. 溶酶体酶的分选是由甘露糖 6-磷酸受体 (M6PR) 进行的，该受体在反式高尔基体网络 (TGN)、核内体和质膜之间转运。在本研究项目中，我们研究了M6PR胞质外结构域在其细胞内运输中的功能，并得到以下结论。（1）GFP-M6PR与氯喹的延时成像和FRAP（光漂白后荧光恢复）分析治疗表明，M6PR 的胞质外结构域是与内吞区室紧密相互作用并被内吞区室保留所必需的。(2) 使用药物 U18666A 进行的实验，以及来自 I 细胞疾病患者的成纤维细胞表明与 M6P 配体的结合参与了尚未确定的 M6PR 转运步骤。 U18666A 可能是研究 M6PR.2 新运输步骤的有用工具。为了研究调节 M6PR 运输的网格蛋白接头 GGA 的功能，我们对 GGA2 进行了 RNAi 和显性失活实验。虽然单独消耗 GGA2 不会导致 M6PR 明显重新分布，但溶酶体酶组织蛋白酶 D 明显错误分泌。这些结果表明，溶酶体酶的错误分选并不总是伴随着剧烈的 M6PR 重新分布。我们将详细分析哪些运输步骤受 GGA 监管。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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。

DOI：
发表时间：
2006
期刊：
Neurosci Lett 410(2)
影响因子：
0
作者：
Yamada S;Isojima Y;Kanamori S;Waguri S;Uchiyama Y;Nagai K.
通讯作者：
Nagai K.

The luminal domain participates in the endosomal trafficking of the cation-independent mannose 6-phosphate receptor.

DOI：
10.1016/j.yexcr.2006.09.024
发表时间：
2006-12
期刊：
Experimental cell research
影响因子：
3.7
作者：
S. Waguri;Yuji Tomiyama;H. Ikeda;T. Hida;N. Sakai;M. Taniike;S. Ebisu;Y. Uchiyama
通讯作者：
S. Waguri;Yuji Tomiyama;H. Ikeda;T. Hida;N. Sakai;M. Taniike;S. Ebisu;Y. Uchiyama

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
影响因子：
0
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