Kwak S P, Hakes D J, Martell K J, Dixon J E
Department of Biological Chemistry, University of Michigan, Ann Arbor 48109-0606.
J Biol Chem. 1994 Feb 4;269(5):3596-604.
Vaccinia phosphatase VH-1 and its mammalian counterparts, including protein-tyrosine phosphatases (PTPase) CL100 and VHR, constitute a novel subfamily of protein-tyrosine phosphatases that exhibits dual substrate specificity for phosphotyrosine- and phosphoserine/threonine-containing substrates. The expression of human VH-1-like PTPase CL100 is rapidly inducible by mitogen stimulation and oxidative stress, suggesting that this gene is transcriptionally regulated. In order to study the mechanism underlying this transcriptional regulation, we isolated the first human gene of this subfamily, the CL100 gene, and characterized its promoter. The gene consists of four exons intervened by three short introns 400-500 base pairs in length. Analysis of the protein sequence encoded by each exon revealed that there is a second region of similarity between CL100 protein and cdc25 in addition to the PTPase catalytic domain. Promoter analysis of the CL100 gene indicates that an 800-base pair region flanking the transcriptional initiation site is sufficient to confer a transcriptional response to serum and 12-O-tetradecanoylphorbol-13-acetate stimulation. The CL100 gene is expressed in numerous tissues, including nonmitotic cells in the brain. Within the brain, CL100 mRNA is localized in discrete neuronal populations, suggesting that this PTPase is likely to play a key role in neurotransmission as well as in mitotic signaling. Finally, although extracellular signal-regulated kinase has recently been shown to act as substrate for CL100 in vitro, we find no clear correspondence between the distribution of extracellular signal-regulated kinase and CL100 mRNA in the brain. The potential significance of a second cdc25 homology domain of CL100 is discussed.
痘苗磷酸酶VH-1及其哺乳动物对应物,包括蛋白酪氨酸磷酸酶(PTPase)CL100和VHR,构成了一个新型的蛋白酪氨酸磷酸酶亚家族,该亚家族对含磷酸酪氨酸和磷酸丝氨酸/苏氨酸的底物具有双重底物特异性。人VH-1样PTPase CL100的表达可被丝裂原刺激和氧化应激迅速诱导,这表明该基因受到转录调控。为了研究这种转录调控的潜在机制,我们分离了该亚家族的首个人类基因CL100基因,并对其启动子进行了表征。该基因由四个外显子组成,中间间隔着三个长度为400 - 500个碱基对的短内含子。对每个外显子编码的蛋白质序列进行分析发现,除了PTPase催化结构域外,CL100蛋白与cdc25之间还有第二个相似区域。对CL100基因的启动子分析表明,转录起始位点两侧一个800个碱基对的区域足以赋予对血清和12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯刺激的转录反应。CL100基因在众多组织中表达,包括大脑中的非有丝分裂细胞。在大脑中,CL100 mRNA定位于离散的神经元群体中,这表明这种PTPase可能在神经传递以及有丝分裂信号传导中起关键作用。最后,尽管最近已证明细胞外信号调节激酶在体外可作为CL100的底物,但我们发现大脑中细胞外信号调节激酶的分布与CL100 mRNA之间没有明显的对应关系。文中还讨论了CL100第二个cdc25同源结构域的潜在意义。