A conserved WXXE motif is an apical delivery determinant of ABC transporter C subfamily isoforms.

IF 2 4区 生物学 Q4 CELL BIOLOGY
Cell structure and function Pub Date : 2023-03-09 Epub Date: 2023-01-25 DOI:10.1247/csf.22049
Md Shajedul Haque, Yoshikazu Emi, Masao Sakaguchi
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引用次数: 0

Abstract

ATP-binding cassette transporter isoform C7 (ABCC7), also designated as cystic fibrosis transmembrane conductance regulator (CFTR), is exclusively targeted to the apical plasma membrane of polarized epithelial cells. Although the apical localization of ABCC7 in epithelia is crucial for the Cl- excretion into lumens, the mechanism regulating its apical localization is poorly understood. In the present study, an apical localization determinant was identified in the N-terminal 80-amino acid long cytoplasmic region of ABCC7 (NT80). In HepG2 cells, overexpression of NT80 significantly disturbed the apical expression of ABCC7 in a competitive manner, suggesting the presence of a sorting determinant in this region. Deletion analysis identified a potential sorting information within a 20-amino acid long peptide (aa 41-60) of NT80. Alanine scanning mutagenesis of this region in full-length ABCC7 further narrowed down the apical localization determinant to four amino acids, W57DRE60. This WDRE sequence was conserved among vertebrate ABCC7 orthologs. Site-directed mutagenesis showed that W57 and E60 were critical for the apical expression of ABCC7, confirming a novel apical sorting determinant of ABCC7. Furthermore, a WXXE motif (tryptophan and glutamic acid residues with two-amino acid spacing) was found to be conserved among the N-terminal regions of apically localized ABCC members with 12-TM configuration. The significance of the WXXE motif was demonstrated for proper trafficking of ABCC4 to the apical plasma membrane.Key words: apical plasma membrane, sorting, ATP-binding cassette transporter, CFTR, MRP4.

保守的 WXXE 基序是 ABC 转运体 C 亚家族同种异构体的顶端输送决定因素。
atp结合盒转运蛋白异构体C7 (ABCC7),也被称为囊性纤维化跨膜传导调节剂(CFTR),专门针对极化上皮细胞的顶质膜。尽管ABCC7在上皮中的顶端定位对Cl-排泄到管腔中至关重要,但其顶端定位的调节机制尚不清楚。在本研究中,在ABCC7 (NT80)的n端80个氨基酸长的细胞质区发现了一个顶端定位决定因子。在HepG2细胞中,NT80的过表达以竞争的方式显著干扰了ABCC7的顶端表达,表明该区域存在分选决定因素。缺失分析在NT80的20个氨基酸长肽(aa 41-60)中发现了潜在的分选信息。对全长ABCC7的这一区域进行丙氨酸扫描诱变,进一步将顶端定位决定因素缩小到4个氨基酸W57DRE60。该WDRE序列在脊椎动物ABCC7同源物中具有保守性。定点突变表明,W57和E60对ABCC7的根尖表达至关重要,证实了ABCC7根尖分选的新决定因素。此外,一个WXXE基序(色氨酸和谷氨酸残基,间隔两个氨基酸)在顶端定位的12-TM结构ABCC成员的n端区域中被发现是保守的。WXXE基序对于ABCC4向根尖质膜的适当转运具有重要意义。关键词:顶质膜,分选,atp结合盒转运体,CFTR, MRP4
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Cell structure and function
Cell structure and function 生物-细胞生物学
CiteScore
2.50
自引率
0.00%
发文量
6
审稿时长
>12 weeks
期刊介绍: Cell Structure and Function is a fully peer-reviewed, fully Open Access journal. As the official English-language journal of the Japan Society for Cell Biology, it is published continuously online and biannually in print. Cell Structure and Function publishes important, original contributions in all areas of molecular and cell biology. The journal welcomes the submission of manuscripts on research areas such as the cell nucleus, chromosomes, and gene expression; the cytoskeleton and cell motility; cell adhesion and the extracellular matrix; cell growth, differentiation and death; signal transduction; the protein life cycle; membrane traffic; and organelles.
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