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磷酸化的Rab11-FIP2与Eps15的相互作用调节顶端连接复合体的组成。

Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition.

作者信息

Lapierre Lynne A, Manning Elizabeth H, Mitchell Kenya M, Caldwell Cathy M, Goldenring James R

机构信息

Section of Surgical Sciences, Vanderbilt University School of Medicine, Nashville, TN 37232.

Epithelial Biology Center, Vanderbilt University School of Medicine, Nashville, TN 37232.

出版信息

Mol Biol Cell. 2017 Apr 15;28(8):1088-1100. doi: 10.1091/mbc.E16-04-0214. Epub 2017 Feb 22.

DOI:10.1091/mbc.E16-04-0214
PMID:28228550
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5391185/
Abstract

MARK2 regulates the establishment of polarity in Madin-Darby canine kidney (MDCK) cells in part through phosphorylation of serine 227 of Rab11-FIP2. We identified Eps15 as an interacting partner of phospho-S227-Rab11-FIP2 (pS227-FIP2). During recovery from low calcium, Eps15 localized to the lateral membrane before pS227-FIP2 arrival. Later in recovery, Eps15 and pS227-FIP2 colocalized at the lateral membrane. In MDCK cells expressing the pseudophosphorylated FIP2 mutant FIP2(S227E), during recovery from low calcium, Eps15 was trapped and never localized to the lateral membrane. Mutation of any of the three NPF domains within GFP-FIP2(S227E) rescued Eps15 localization at the lateral membrane and reestablished single-lumen cyst formation in GFP-FIP2(S227E)-expressing cells in three-dimensional (3D) culture. Whereas expression of GFP-FIP2(S227E) induced the loss of E-cadherin and occludin, mutation of any of the NPF domains of GFP-FIP2(S227E) reestablished both proteins at the apical junctions. Knockdown of Eps15 altered the spatial and temporal localization of pS227-FIP2 and also elicited formation of multiple lumens in MDCK 3D cysts. Thus an interaction of Eps15 and pS227-FIP2 at the appropriate time and location in polarizing cells is necessary for proper establishment of epithelial polarity.

摘要

MARK2部分通过对Rab11 - FIP2的丝氨酸227进行磷酸化来调节Madin - Darby犬肾（MDCK）细胞极性的建立。我们鉴定出Eps15是磷酸化的S227 - Rab11 - FIP2（pS227 - FIP2）的相互作用蛋白。在从低钙环境恢复的过程中，Eps15在pS227 - FIP2到达之前定位于侧膜。在恢复后期，Eps15和pS227 - FIP2在侧膜共定位。在表达假磷酸化FIP2突变体FIP2(S227E)的MDCK细胞中，从低钙环境恢复时，Eps15被困住且从未定位于侧膜。GFP - FIP2(S227E)内三个NPF结构域中的任何一个发生突变，都能挽救Eps15在侧膜的定位，并在三维（3D）培养的表达GFP - FIP2(S227E)的细胞中重新建立单腔囊肿的形成。虽然GFP - FIP2(S227E)的表达导致E - 钙黏蛋白和闭合蛋白的丢失，但GFP - FIP2(S227E)的任何一个NPF结构域发生突变，都能使这两种蛋白重新定位于顶端连接。敲低Eps15会改变pS227 - FIP2的时空定位，也会引发MDCK 3D囊肿中多个腔的形成。因此，在极化细胞中，Eps15和pS227 - FIP2在适当的时间和位置相互作用，对于上皮极性的正确建立是必要的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/b76baec59576/1088fig10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/134bfd32b8de/1088fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/8c5a9539c355/1088fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/ae87611a0f56/1088fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/bcc3a85ee5b1/1088fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/aa8c30ade97f/1088fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/4d230e0c4df6/1088fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/d868d30bb37f/1088fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/cc546f909289/1088fig8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/b21c3c0fa9d1/1088fig9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/b76baec59576/1088fig10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/134bfd32b8de/1088fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/8c5a9539c355/1088fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/ae87611a0f56/1088fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/bcc3a85ee5b1/1088fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/aa8c30ade97f/1088fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/4d230e0c4df6/1088fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/d868d30bb37f/1088fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/cc546f909289/1088fig8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/b21c3c0fa9d1/1088fig9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9230/5391185/b76baec59576/1088fig10.jpg

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