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秀丽隐杆线虫肠道半胱氨酸蛋白酶基因cpr-1的调控:对GATA基序的需求

Regulation of the Caenorhabditis elegans gut cysteine protease gene cpr-1: requirement for GATA motifs.

作者信息

Britton C, McKerrow J H, Johnstone I L

机构信息

Wellcome Unit of Molecular Parasitology, University of Glasgow, Anderson College, 56 Dumbarton Road, Glasgow, G11 6NU, UK.

出版信息

J Mol Biol. 1998;283(1):15-27. doi: 10.1006/jmbi.1998.2093.

DOI:10.1006/jmbi.1998.2093
PMID:9761670
Abstract

Expression of the Caenorhabditis elegans cysteine protease gene cpr-1 is regulated both spatially and temporally. In situ hybridisation and Northern blot analysis have shown that this gene is expressed exclusively in gut cells of all developmental stages except the embryo. We now show by transgenic transformation with cpr-1/lac Z reporter gene constructs that a sequence contained within the cpr-1 5' flanking region can direct this spatial and temporal expression. Deletion analysis of the cpr-1 promoter indicates that as little as 212 bp of upstream sequence is sufficient for this expression, although more upstream sequence may be involved in quantitative regulation of expression. Mutation of two GATA-like sequence elements at positions -51 and -147 upstream of the transcription start site ablates all expression, indicating an essential role in cpr-1 regulation. A concatemer of the cpr-1 -147 GATA motif placed upstream of minimal promoter/lac Z reporter gene constructs results in strong reporter gene expression in gut cells of larval stages and also in embryos. Weak expression is also detected in hypodermal cells. This pattern is reversed in the adult stage with strong expression in hypodermal cells and weaker expression in gut cells. Our findings suggest that spatial and temporal regulation of the cpr-1 gene is complex and involves activation by a GATA-like transcription factor.

摘要

秀丽隐杆线虫半胱氨酸蛋白酶基因cpr-1的表达在空间和时间上均受到调控。原位杂交和Northern印迹分析表明,该基因仅在除胚胎外的所有发育阶段的肠道细胞中表达。我们现在通过用cpr-1/lac Z报告基因构建体进行转基因转化表明,cpr-1 5'侧翼区域内包含的一个序列可以指导这种时空表达。对cpr-1启动子的缺失分析表明,尽管更多的上游序列可能参与表达的定量调控,但仅212 bp的上游序列就足以实现这种表达。转录起始位点上游-51和-147位置的两个GATA样序列元件的突变消除了所有表达,表明其在cpr-1调控中起重要作用。将cpr-1 -147 GATA基序的串联体置于最小启动子/lac Z报告基因构建体的上游,导致幼虫阶段的肠道细胞以及胚胎中报告基因的强烈表达。在皮下细胞中也检测到弱表达。在成虫阶段,这种模式相反,皮下细胞中表达强烈,肠道细胞中表达较弱。我们的研究结果表明,cpr-1基因的时空调控很复杂,涉及一种GATA样转录因子的激活。

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