Muratake T, Hayashi S, Ichimura Y, Morii K, Kuwano R, Ichikawa T, Kumanishi T, Isobe T, Watanabe M, Kondo H
National Saigata Hospital, Niigata Prefecture, Japan.
Mol Neurobiol. 1995 Aug-Dec;11(1-3):223-30. doi: 10.1007/BF02740697.
14.3.3 protein, a brain-specific protein, is an activator of tyrosine and tryptophan hydroxylases, key enzymes for biosynthesis of dopamine and serotonin. In this article, we describe cloning of cDNA for human brain 14.3.3 eta chain and expression of 14.3.3 eta chain mRNA in some human cultured cells. The cloned cDNA is 1730 bp long and contains 191 bp of a 5'-noncoding region, the complete 738 bp of coding region, and 801 bp of a 3'-noncoding region, containing three polyadenylation signals. This cDNA encoded a polypeptide of 246 amino acids (M(r) 28,196). Furthermore, using in situ hybridization histochemistry, the expression of mRNA for this protein was examined in the rat central nervous system. In situ hybridization histochemistry indicated that 14.3.3 eta chain mRNA is detected not only in the monoamine-synthetic neurons, but also in other neurons in the discrete nuclei, which synthesize neither cathecholamine nor serotonin. Northern blot analysis demonstrated that the addition of methamphetamine into the cultured medium increased the mRNA level for 14.3.3 eta chain in U-251 cells, but did not increase that of GFAP.
14.3.3蛋白是一种脑特异性蛋白，是酪氨酸和色氨酸羟化酶的激活剂，这两种酶是多巴胺和5-羟色胺生物合成的关键酶。在本文中，我们描述了人脑14.3.3 η链cDNA的克隆以及14.3.3 η链mRNA在一些人培养细胞中的表达。克隆的cDNA长1730 bp，包含191 bp的5'非编码区、完整的738 bp编码区和801 bp的3'非编码区，含有三个聚腺苷酸化信号。该cDNA编码一个由246个氨基酸组成的多肽（分子量28,196）。此外，利用原位杂交组织化学技术，在大鼠中枢神经系统中检测了该蛋白mRNA的表达。原位杂交组织化学表明，不仅在单胺合成神经元中能检测到14.3.3 η链mRNA，而且在离散核团中既不合成儿茶酚胺也不合成5-羟色胺的其他神经元中也能检测到。Northern印迹分析表明，向培养基中添加甲基苯丙胺可增加U-251细胞中14.3.3 η链的mRNA水平，但不会增加GFAP的mRNA水平。
