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Cx31 is assembled and trafficked to cell surface by ER-Golgi pathway and degraded by proteasomal or lysosomal pathways

查看全文 作  者:[2]LiQiangHE;[1]FangCAI;[1]YuLIU;[1]MuJunLIU;[1]ZhiPingTAN;[1]QianPAN;[1]FaiYanFANG;[1]DeShengLIANG;[1]LingQianWU;[1]ZhiGaoLONG;[1]HePingDAI;[1]KunXIA;[1]JiaHuiXIA;[1]ZhuoHuaZHANG 高影响力作者 机构地区:[1]NationalLaboratoryofMedicalGenetics,CentralSouthUniversity,Changsha410078,China.;[2]InstituteofMedicalGenetics,MedicalSchoolofShang-haiJiaoTongUniversity.24#,lane1400,WestBeijingRoad,Shanghai,China.高影响力机构 出  处:《Cell Research》索引2005年第15卷第6期,共10页高影响力期刊 基  金:supported by 'the National High Tech-nology Research and Development Program of China, No.2002BA711A07-03, 08;the Major State Basic ResearchDevelopment Program of China, No. 2001CB510302 and2004CB518800;the National Natural Science Foun-dation of China, No. 30370737. 摘  要:Gap junctions, consisting of connexins, allow the exchange of small molecules (<1 kD) between adjacent cells, thusproviding a mechanism for synchronizing the responses of groups of cells to environmental stimuli. Connexin 31 is amember of the connexin family. Mutations on connexin 31 are associated with erythrokeratodermia variabilis, hearingimpairment and peripheral neuropathy. However, the pathological mechanism for connexin 31 mutants in these diseasesare still unknown. In this study, we analyzed the assembly, trafficking and metabolism of connexin 31 in HeLa cellsstably expressing connexin 31. Calcein transfer assay showed that calcein transfer was inhibited when cells weretreated with Brefeldin A or cytochalasin D, but not when treated with nocodazole or α-glycyrrhetinic acid, suggestingthat Golgi apparatus and actin filaments, but not microtubules, are crucial to the trafficking and assembly of connexin31, as well as the formation of gap junction intercellular communication by connexin 31. Additionally, α-glycyrrhetinicacid did not effectively inhibit gap junctional intercellular communication formed by connexin 31. Pulse-chase assayrevealed that connexin 31 had a half-life of about 6 h. Moreover, Western blotting and fluorescent staining demonstratedthat in HeLa cells stably expressing connexin 31, the amount of connexin 31 was significantly increased after these cellswere treated with proteasomal or lysosomal inhibitors. These findings indicate that connexin 31 was rapidly renewed,and possibly degraded by both proteasomal and lysosomal pathways. 关 键 词:细胞 连接蛋白 Cx31 新陈代谢 运输路径 蛋白酶 溶菌酶
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