STRUCTURE, MAPPING, AND EXPRESSION OF ERP, A GROWTH FACTOR-INDUCIBLE GENE ENCODING A NONTRANSMEMBRANE PROTEIN-TYROSINE-PHOSPHATASE, AND EFFECT OF ERP ON CELL-GROWTH
STRUCTURE, MAPPING, AND EXPRESSION OF ERP, A GROWTH FACTOR-INDUCIBLE GENE ENCODING A NONTRANSMEMBRANE PROTEIN-TYROSINE-PHOSPHATASE, AND EFFECT OF ERP ON CELL-GROWTH
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DOI：
10.1128/mcb.13.9.5195
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发表时间：
1993-09-01
影响因子：
5.3
通讯作者：
BRAVO, R
中科院分区：
文献类型：
--
作者：
NOGUCHI, T;METZ, R;BRAVO, R
我们的特点是生长因子诱导基因，erp，并证明它编码一个367个氨基酸的非跨膜酪氨酸磷酸酶蛋白具有显着的相似性的牛痘病毒HI蛋白。免疫沉淀分析表明，erp蛋白，ERP，是快速诱导静止成纤维细胞的血清刺激后。ERP已被表达为与谷胱甘肽S-转移酶的融合蛋白，并显示具有酪氨酸和丝氨酸蛋白磷酸酶活性。ERP的酶活性取决于还原剂如二硫苏糖醇的存在，其酪氨酸磷酸酶活性被钒酸钠抑制，钒酸钠是蛋白酪氨酸磷酸酶的有效抑制剂。用表达完整ERP蛋白的载体转染后获得的稳定NIH 3 T3克隆的数量与用表达突变的失活ERP蛋白的载体转染后相比减少了90%以上。剩余的ERP表达克隆呈现出双核和多核细胞比例的显著增加和增殖率的降低。对基因组结构的研究表明erp转录单位长度为2.8kbp，分为4个外显子。erp基因定位于小鼠基因组的17 A2 - 17 C区域。我们的研究结果表明，即刻早期基因erp的蛋白产物对细胞增殖有负面影响。
We have characterized a growth factor-inducible gene, erp, and demonstrated that it encodes a 367-amino-acid nontransmembrane tyrosine phosphatase protein with significant similarity to the vaccinia virus HI protein. Immunoprecipitation analyses show that the erp protein, ERP, is rapidly induced following serum stimulation of quiescent fibroblasts. ERP has been expressed as a fusion protein with glutathione S-transferase and shown to have tyrosine as well as serine protein phosphatase activity. The enzymatic activity of ERP depends on the presence of reducing agents such as dithiothreitol, and its tyrosine phosphatase activity is inhibited by sodium vanadate, a potent inhibitor of protein tyrosine phosphatases. The number of stable NIH 3T3 clones obtained after transfection with a vector expressing the complete ERP protein is reduced more than 90% compared with that after transfection with a vector expressing a mutated inactive ERP protein. The remaining ERP-expressing clones present a significant increase in the proportion of bi- and multinucleated cells and a decrease in proliferation rate. Studies on the genomic structure reveal that the erp transcription unit is 2.8 kbp long and split into four exons. The erp gene maps to the 17A2-17C region of the murine genome. Our results demonstrate that the protein product of the immediate-early gene erp has a negative effect on cell proliferation.