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尿路上皮蛋白通过高尔基体诱导其片段化：新型体外模型的新见解。

Uroplakin traffic through the Golgi apparatus induces its fragmentation: new insights from novel in vitro models.

机构信息

Institute of Cell Biology, Faculty of Medicine, University of Ljubljana, Vrazov trg 2, SI-1000, Ljubljana, Slovenia.

Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078, Pozzuoli, (NA), Italy.

出版信息

Sci Rep. 2017 Oct 9;7(1):12842. doi: 10.1038/s41598-017-13103-x.

DOI:10.1038/s41598-017-13103-x
PMID:28993693
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5634464/
Abstract

Uroplakins (UPs) play an essential role in maintaining an effective urothelial permeability barrier at the level of superficial urothelial cell (UC) layer. Although the organization of UPs in the apical plasma membrane (PM) of UCs is well known, their transport in UCs is only partially understood. Here, we dissected trafficking of UPs and its differentiation-dependent impact on Golgi apparatus (GA) architecture. We demonstrated that individual subunits UPIb and UPIIIa are capable of trafficking from the endoplasmic reticulum to the GA in UCs. Moreover, UPIb, UPIIIa or UPIb/UPIIIa expressing UCs revealed fragmentation and peripheral redistribution of Golgi-units. Notably, expression of UPIb or UPIb/UPIIIa triggered similar GA fragmentation in MDCK and HeLa cells that do not express UPs endogenously. The colocalization analysis of UPIb/UPIIIa-EGFP and COPI, COPII or clathrin suggested that UPs follow constitutively the post-Golgi route to the apical PM. Depolymerisation of microtubules leads to complete blockade of the UPIb/UPIIIa-EGFP post-Golgi transport, while disassembly of actin filaments shows significantly reduced delivery of UPIb/UPIIIa-EGFP to the PM. Our findings show the significant effect of the UPs expression on the GA fragmentation, which enables secretory Golgi-outpost to be distributed as close as possible to the sites of cargo delivery at the PM.

摘要

尿路上皮蛋白（Uroplakins，UPs）在维持浅表尿路上皮细胞（UC）层的有效尿路上皮通透性屏障方面发挥着重要作用。尽管 UPs 在 UC 的顶质膜（PM）中的组织是众所周知的，但它们在 UC 中的转运仅部分被理解。在这里，我们剖析了 UPs 的转运及其对高尔基器（GA）结构的分化依赖性影响。我们证明了单个亚基 UPIb 和 UPIIIa 能够从内质网转运到 UC 的 GA。此外，表达 UPIb、UPIIIa 或 UPIb/UPIIIa 的 UC 显示出高尔基体单元的碎片化和外周再分布。值得注意的是，在不表达内源性 UPs 的 MDCK 和 HeLa 细胞中，表达 UPIb 或 UPIb/UPIIIa 会引发类似的 GA 碎片化。UPIb/UPIIIa-EGFP 与 COPI、COPII 或网格蛋白的共定位分析表明，UPs 遵循组成型的高尔基后途径到达顶质膜。微管的去聚合导致 UPIb/UPIIIa-EGFP 的高尔基后转运完全阻断，而肌动蛋白丝的解体显示 UPIb/UPIIIa-EGFP 向 PM 的递送显著减少。我们的研究结果表明，UPs 的表达对 GA 碎片化有显著影响，这使得分泌型高尔基前哨能够尽可能靠近 PM 处的货物输送位点分布。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/574af5ed9a83/41598_2017_13103_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/bc9f0b1a34ca/41598_2017_13103_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/a5fa79a206b3/41598_2017_13103_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/b74ded6f40a9/41598_2017_13103_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/6e5921ded028/41598_2017_13103_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/06966fda910c/41598_2017_13103_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/b99a11563176/41598_2017_13103_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/574af5ed9a83/41598_2017_13103_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/bc9f0b1a34ca/41598_2017_13103_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/a5fa79a206b3/41598_2017_13103_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/b74ded6f40a9/41598_2017_13103_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/6e5921ded028/41598_2017_13103_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/06966fda910c/41598_2017_13103_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/b99a11563176/41598_2017_13103_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b40/5634464/574af5ed9a83/41598_2017_13103_Fig7_HTML.jpg

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