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ELT-2,一种来自线虫秀丽隐杆线虫的第二个GATA因子。

elt-2, a second GATA factor from the nematode Caenorhabditis elegans.

作者信息

Hawkins M G, McGhee J D

机构信息

Department of Medical Biochemistry, University of Calgary, Alberta, Canada.

出版信息

J Biol Chem. 1995 Jun 16;270(24):14666-71. doi: 10.1074/jbc.270.24.14666.

Abstract

We have previously shown that a tandem pair of (A/T)GATA(A/G) sequences in the promoter region of the Caenorhabditis elegans gut esterase gene (ges-1) controls the tissue specificity of ges-1 expression in vivo. The ges-1 GATA region was used as a probe to screen a C. elegans cDNA expression library, and a gene for a new C. elegans GATA-factor (named elt-2) was isolated. The longest open reading frame in the elt-2 cDNA codes for a protein of M(r) 47,000 with a single zinc finger domain, similar (approximately 75% amino acid identity) to the C-terminal fingers of all other two-fingered GATA factors isolated to date. A similar degree of relatedness is found with the single-finger DNA binding domains of GATA factors identified in invertebrates. An upstream region in the ELT-2 protein with the sequence C-X2-C-X16-C-X2-C has some of the characteristics of a zinc finger domain but is highly diverged from the zinc finger domains of other GATA factors. The elt-2 gene is expressed as an SL1 trans-spliced message, which can be detected at all stages of development except oocytes; however, elt-2 message levels are 5-10-fold higher in embryos than in other stages. The genomic clone for elt-2 has been characterized and mapped near the center of the C. elegans X chromosome, ELT-2 protein, produced by in vitro transcription-translation, binds to ges-1 GATA-containing oligonucleotides similar to a factor previously identified in C. elegans embryo extracts, both as assayed by electrophoretic migration and by competition with wild type and mutant oligonucleotides. However, there is as yet no direct evidence that elt-2 does or does not control ges-1.

摘要

我们之前已经表明,秀丽隐杆线虫肠道酯酶基因(ges-1)启动子区域中的一对串联(A/T)GATA(A/G)序列在体内控制ges-1表达的组织特异性。ges-1 GATA区域用作探针筛选秀丽隐杆线虫cDNA表达文库,并分离出一个新的秀丽隐杆线虫GATA因子(命名为elt-2)的基因。elt-2 cDNA中最长的开放阅读框编码一个分子量为47,000的蛋白质,具有单个锌指结构域,与迄今为止分离出的所有其他双指GATA因子的C末端指相似(氨基酸同一性约为75%)。在无脊椎动物中鉴定出的GATA因子的单指DNA结合结构域也发现了类似程度的相关性。ELT-2蛋白中具有C-X2-C-X16-C-X2-C序列的上游区域具有一些锌指结构域的特征,但与其他GATA因子的锌指结构域高度不同。elt-2基因表达为SL1反式剪接的信息,除卵母细胞外,在发育的所有阶段都可检测到;然而,elt-2信息水平在胚胎中比在其他阶段高5至10倍。elt-2的基因组克隆已被表征并定位在秀丽隐杆线虫X染色体的中心附近,通过体外转录-翻译产生的ELT-2蛋白与含有ges-1 GATA的寡核苷酸结合,类似于先前在秀丽隐杆线虫胚胎提取物中鉴定的一种因子,这通过电泳迁移和与野生型及突变寡核苷酸的竞争来测定。然而,尚无直接证据表明elt-2是否控制ges-1。

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