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Connexin 31.1 degradation requires the Clathrin-mediated autophagy in NSCLC cell H1299

Connexins have relative short half-lives. Connexin 31.1 (Cx31.1) was newly reported to be down-regulated in non-small cell lung cancer cell lines, and displayed tumour-suppressive properties. However, no reports describing how a cell regulates Cx31.1 level were found. In this study, Cx31.1 was sugge...

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Autores principales: Zhu, Xingli, Ruan, Zhenchao, Yang, Xiufang, Chu, Kaili, Wu, Hai, Li, Yao, Huang, Yan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Blackwell Publishing Ltd 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4288368/
https://www.ncbi.nlm.nih.gov/pubmed/25388970
http://dx.doi.org/10.1111/jcmm.12470
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author Zhu, Xingli
Ruan, Zhenchao
Yang, Xiufang
Chu, Kaili
Wu, Hai
Li, Yao
Huang, Yan
author_facet Zhu, Xingli
Ruan, Zhenchao
Yang, Xiufang
Chu, Kaili
Wu, Hai
Li, Yao
Huang, Yan
author_sort Zhu, Xingli
collection PubMed
description Connexins have relative short half-lives. Connexin 31.1 (Cx31.1) was newly reported to be down-regulated in non-small cell lung cancer cell lines, and displayed tumour-suppressive properties. However, no reports describing how a cell regulates Cx31.1 level were found. In this study, Cx31.1 was suggested to be degraded through both ubiquitin–proteasome system (UPS) and autophagy. Blockage of UPS with MG-132 increased Cx31.1 level, but could not inhibit the degradation of Cx31.1 completely. In H1299 cells stably expressing Cx31.1, Cx31.1 reduced when autophagy was induced through starvation or Brefeldin A treatment. Knockdown of autophagy-related protein ATG5 could increase the cellular level of Cx31.1 both under normal growth condition and starvation-induced autophagy. Colocalization of Cx31.1 and autophagy marker light chain 3 (LC3) was revealed by immunofluorescence analysis. Coimmunoprecipitation and immunofluorescence showed that Cx31.1 might interact with clathrin heavy chain which was newly reported to regulate autophagic lysosome reformation (ALR) and controls lysosome homoeostasis. When clathrin expression was knockdown by siRNA treatment, the level of Cx31.1 increased prominently both under normal growth condition and starvation-induced autophagy. Under starvation-induced autophagy, LC3-II levels were slightly accumulated with siCla. treatment compared to that of siNC, which could be ascribed to that clathrin knockdown impaired the late stage of autophagy, ALR. Taken together, we found autophagy contributed to Cx31.1 degradation, and clathrin might be involved in the autophagy of Cx31.1.
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spelling pubmed-42883682015-01-21 Connexin 31.1 degradation requires the Clathrin-mediated autophagy in NSCLC cell H1299 Zhu, Xingli Ruan, Zhenchao Yang, Xiufang Chu, Kaili Wu, Hai Li, Yao Huang, Yan J Cell Mol Med Original Articles Connexins have relative short half-lives. Connexin 31.1 (Cx31.1) was newly reported to be down-regulated in non-small cell lung cancer cell lines, and displayed tumour-suppressive properties. However, no reports describing how a cell regulates Cx31.1 level were found. In this study, Cx31.1 was suggested to be degraded through both ubiquitin–proteasome system (UPS) and autophagy. Blockage of UPS with MG-132 increased Cx31.1 level, but could not inhibit the degradation of Cx31.1 completely. In H1299 cells stably expressing Cx31.1, Cx31.1 reduced when autophagy was induced through starvation or Brefeldin A treatment. Knockdown of autophagy-related protein ATG5 could increase the cellular level of Cx31.1 both under normal growth condition and starvation-induced autophagy. Colocalization of Cx31.1 and autophagy marker light chain 3 (LC3) was revealed by immunofluorescence analysis. Coimmunoprecipitation and immunofluorescence showed that Cx31.1 might interact with clathrin heavy chain which was newly reported to regulate autophagic lysosome reformation (ALR) and controls lysosome homoeostasis. When clathrin expression was knockdown by siRNA treatment, the level of Cx31.1 increased prominently both under normal growth condition and starvation-induced autophagy. Under starvation-induced autophagy, LC3-II levels were slightly accumulated with siCla. treatment compared to that of siNC, which could be ascribed to that clathrin knockdown impaired the late stage of autophagy, ALR. Taken together, we found autophagy contributed to Cx31.1 degradation, and clathrin might be involved in the autophagy of Cx31.1. Blackwell Publishing Ltd 2015-01 2014-11-11 /pmc/articles/PMC4288368/ /pubmed/25388970 http://dx.doi.org/10.1111/jcmm.12470 Text en © 2014 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Zhu, Xingli
Ruan, Zhenchao
Yang, Xiufang
Chu, Kaili
Wu, Hai
Li, Yao
Huang, Yan
Connexin 31.1 degradation requires the Clathrin-mediated autophagy in NSCLC cell H1299
title Connexin 31.1 degradation requires the Clathrin-mediated autophagy in NSCLC cell H1299
title_full Connexin 31.1 degradation requires the Clathrin-mediated autophagy in NSCLC cell H1299
title_fullStr Connexin 31.1 degradation requires the Clathrin-mediated autophagy in NSCLC cell H1299
title_full_unstemmed Connexin 31.1 degradation requires the Clathrin-mediated autophagy in NSCLC cell H1299
title_short Connexin 31.1 degradation requires the Clathrin-mediated autophagy in NSCLC cell H1299
title_sort connexin 31.1 degradation requires the clathrin-mediated autophagy in nsclc cell h1299
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4288368/
https://www.ncbi.nlm.nih.gov/pubmed/25388970
http://dx.doi.org/10.1111/jcmm.12470
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