pH-mediated upregulation of AQP1 gene expression through the Spi-B transcription factor

IF 2.946 Q3 Biochemistry, Genetics and Molecular Biology BMC Molecular Biology Pub Date : 2018-03-20 DOI:10.1186/s12867-018-0104-9
Yihui Zhai, Hong Xu, Qian Shen, Franz Schaefer, Claus P. Schmitt, Jing Chen, Haimei Liu, Jialu Liu, Jiaojiao Liu
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引用次数: 3

Abstract

Bicarbonate-based peritoneal dialysis (PD) fluids enhance the migratory capacity and damage-repair ability of human peritoneal mesothelial cells by upregulating AQP1. However, little is known about the underlying molecular mechanisms.

Here we used HEK-293T cells to investigate the effect of pH on AQP1 gene transcription levels. We found that AQP1 mRNA levels increases with pH. Transfection of HEK-293T cells with luciferase reporter vectors containing different regions of the AQP1 promoter identified an upstream region in the AQP1 gene between ??2200 and –?2300?bp as an enhancer required for pH-mediated regulation of AQP1 expression. Site-directed mutagenesis of this specific promoter region revealed a critical region between ??2257 and ??2251?bp, and gene knock-down experiments and ChIP assays suggested that the Spi-B transcription factor SPIB is involved in pH-mediated regulation of AQP1 expression.

We identified an upstream region in the AQP1 gene and the transcription factor SPIB that are critically involved in pH-mediated regulation of AQP1 expression. These findings provide the basis for further studies on the pH- and buffer-dependent effects of PD fluids on peritoneal membrane integrity and function.

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ph通过Spi-B转录因子介导AQP1基因表达上调
碳酸氢盐基腹膜透析(PD)液通过上调AQP1增强人腹膜间皮细胞的迁移能力和损伤修复能力。然而,人们对其潜在的分子机制知之甚少。我们利用HEK-293T细胞研究pH对AQP1基因转录水平的影响。我们发现AQP1 mRNA水平随ph升高。用含有AQP1启动子不同区域的荧光素酶报告载体转染HEK-293T细胞,发现AQP1基因的上游区域位于??2200 - 2300?bp作为ph介导的AQP1表达调节所需的增强子。对这一特定启动子区域的定点突变揭示了在??2257和2251?基因敲除实验和ChIP分析提示SPIB - b转录因子SPIB参与ph介导的AQP1表达调控。我们确定了AQP1基因的上游区域和转录因子SPIB,它们在ph介导的AQP1表达调控中起关键作用。这些发现为进一步研究PD液的pH和缓冲液对腹膜完整性和功能的影响提供了基础。
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来源期刊
BMC Molecular Biology
BMC Molecular Biology 生物-生化与分子生物学
CiteScore
4.80
自引率
0.00%
发文量
0
审稿时长
>12 weeks
期刊介绍: BMC Molecular Biology is an open access journal publishing original peer-reviewed research articles in all aspects of DNA and RNA in a cellular context, encompassing investigations of chromatin, replication, recombination, mutation, repair, transcription, translation and RNA processing and function.
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