Regulation of the Caenorhabditis elegans gut cysteine protease gene cpr-1:: Requirement for GATA motifs
Regulation of the Caenorhabditis elegans gut cysteine protease gene cpr-1:: Requirement for GATA motifs
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DOI：
10.1006/jmbi.1998.2093
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发表时间：
1998-10-16
影响因子：
5.6
通讯作者：
Johnstone, LL
中科院分区：
文献类型：
--
作者：
Britton, C;McKerrow, JH;Johnstone, LL
秀丽线虫半胱氨酸蛋白酶基因CPR-1的表达在空间和时间上都受到调控。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报告基因构建的转基因转化表明，CPR-1 5‘侧翼区内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，尽管可能有更多的上游序列参与表达的定量调控，但仅212bp的上游序列就足以满足这种表达。转录起始点上游-51位和-147位的两个类似GATA的序列元件的突变使所有的表达消失，表明在CPR-L的调控中发挥了重要作用。位于最小启动子/lac Z报告基因上游的CPR-1-147 GATA基序的串联导致报告基因在幼虫阶段的肠道细胞和胚胎中强表达。在皮下细胞中也检测到微弱的表达。这种模式在成虫阶段相反，在皮下细胞中表达较强，在肠道细胞中表达较弱。我们的发现表明，CPR-1基因的时空调控是复杂的，涉及到GATA样转录因子的激活。(C)1998年学术出版社。
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-l 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. (C) 1998 Academic Press.