Spieth J, Shim Y H, Lea K, Conrad R, Blumenthal T
Program in Molecular, Cellular and Developmental Biology, Indiana University, Bloomington 47405.
Mol Cell Biol. 1991 Sep;11(9):4651-9. doi: 10.1128/mcb.11.9.4651-4659.1991.
The short, asymmetrical DNA sequence to which the vertebrate GATA family of transcription factors binds is present in some Caenorhabditis elegans gene regulatory regions: it is required for activation of the vitellogenin genes and is also found just 5' of the TATA boxes of tra-2 and the msp genes. In vertebrates GATA-1 is specific to erythroid lineages, whereas GATA-2 and GATA-3 are present in multiple tissues. In an effort to identify the trans-acting factors that may recognize this sequence element in C. elegans, we used a degenerate oligonucleotide to clone a C. elegans homolog to this gene. We call this gene elt-1 (erythrocytelike transcription factor). It is single copy and specifies a 1.75-kb mRNA that is present predominantly, if not exclusively, in embryos. The region of elt-1 encoding two zinc fingers is remarkably similar to the DNA-binding domain of the vertebrate GATA-binding proteins. However, outside of the DNA-binding domains the amino acid sequences are quite divergent. Nevertheless, introns are located at identical or nearly identical positions in elt-1 and the mouse GATA-1 gene. In addition, elt-1 mRNA is trans-spliced to the 22-base untranslated leader, SL1. The DNA upstream of the elt-1 TATA box contains eight copies of the GATA recognition sequence within the first 300 bp, suggesting that elt-1 may be autogenously regulated. Our results suggest that the specialized role of GATA-1 in erythroid gene expression was derived after separation of the nematodes and the line that led to the vertebrates, since C. elegans lacks an erythroid lineage.
脊椎动物转录因子GATA家族所结合的短的、不对称DNA序列存在于一些秀丽隐杆线虫的基因调控区域:它是卵黄蛋白原基因激活所必需的,并且也在tra-2和msp基因的TATA框的5'端被发现。在脊椎动物中,GATA-1对红细胞谱系具有特异性,而GATA-2和GATA-3存在于多种组织中。为了鉴定可能识别秀丽隐杆线虫中这个序列元件的反式作用因子,我们使用简并寡核苷酸克隆了该基因的秀丽隐杆线虫同源物。我们将这个基因称为elt-1(类红细胞转录因子)。它是单拷贝的,编码一个1.75-kb的mRNA,该mRNA主要(如果不是唯一的话)存在于胚胎中。elt-1编码两个锌指的区域与脊椎动物GATA结合蛋白的DNA结合结构域非常相似。然而,在DNA结合结构域之外,氨基酸序列差异很大。尽管如此,内含子在elt-1和小鼠GATA-1基因中位于相同或几乎相同的位置。此外,elt-1 mRNA被反式剪接到22个碱基的非翻译前导序列SL1。elt-1 TATA框上游的DNA在前300 bp内包含八个GATA识别序列拷贝,这表明elt-1可能是自身调节的。我们的结果表明,由于秀丽隐杆线虫缺乏红细胞谱系,GATA-1在红细胞基因表达中的特殊作用是在线虫和导致脊椎动物的谱系分离之后产生的。