Freedman J H, Slice L W, Dixon D, Fire A, Rubin C S
Department of Molecular Pharmacology, Atran Laboratories, Albert Einstein College of Medicine, Bronx, New York 10461.
J Biol Chem. 1993 Feb 5;268(4):2554-64.
Two genes (mtl-1 and mtl-2) that encode the novel metallothioneins (MTs) of Caenorhabditis elegans (CeMTs) were cloned and characterized. Both genes contain a single intron that interrupts codon 6 and short 3'-untranslated regions. However, their promotor regions are distinctively non-homologous. The mtl-2 promoter contains a TATAA box and a single putative metal regulatory element. These elements are absent in the mtl-1 promoter. Nevertheless, both CeMT1 and CeMT2 mRNAs are induced by cadmium and contain precisely initiated, 5'-untranslated sequences. The inducibility and cell type specificity of metallothionein gene expression were investigated in transgenic C. elegans that carry the lacZ (beta-galactosidase) reporter gene under the control of an mtl-1 or mtl-2 promoter sequence. Upon treatment of transgenic C. elegans with cadmium or heat stress, the mtl-2:lacZ fusion gene is abundantly and exclusively expressed in the intestinal cells of larvae and adult animals. Expression is not detected in the absence of metal or heat shock. In contrast, an mtl-1:lacZ construct is constitutively expressed in the pharynx and induced by cadmium and heat shock in the intestinal cells of C. elegans larvae. The metal-inducible expression of the mtl-1:lacZ gene is attenuated in adult transgenic nematodes. Thus, the activity of each mtl promoter is modulated by metals as well as developmental and environmental factors.
克隆并鉴定了两个编码秀丽隐杆线虫新型金属硫蛋白(CeMTs)的基因(mtl-1和mtl-2)。这两个基因都含有一个打断第6密码子的内含子和短的3'非翻译区。然而,它们的启动子区域明显非同源。mtl-2启动子包含一个TATAA盒和一个单一的假定金属调节元件。这些元件在mtl-1启动子中不存在。尽管如此,CeMT1和CeMT2 mRNA都能被镉诱导,并且含有精确起始的5'非翻译序列。在携带受mtl-1或mtl-2启动子序列控制的lacZ(β-半乳糖苷酶)报告基因的转基因秀丽隐杆线虫中,研究了金属硫蛋白基因表达的诱导性和细胞类型特异性。用镉或热应激处理转基因秀丽隐杆线虫后,mtl-2:lacZ融合基因在幼虫和成年动物的肠道细胞中大量且特异性表达。在没有金属或热休克的情况下未检测到表达。相比之下,mtl-1:lacZ构建体在咽部组成性表达,并在秀丽隐杆线虫幼虫的肠道细胞中被镉和热休克诱导。mtl-1:lacZ基因的金属诱导表达在成年转基因线虫中减弱。因此,每个mtl启动子的活性受到金属以及发育和环境因素的调节。