Isolation and characterization of a cDNA clone that codes for human spermidine/spermine N1-acetyltransferase.
Isolation and characterization of a cDNA clone that codes for human spermidine/spermine N1-acetyltransferase.
复制标题
DOI：
--
复制
发表时间：
1991-01
期刊：
影响因子：
--
通讯作者：
R. Casero;Paul Celano;S. J. Ervin;Nancy B. Applegren;Laurie Wiest;Anthony E. Pegg
中科院分区：
文献类型：
--
作者：
R. Casero;Paul Celano;S. J. Ervin;Nancy B. Applegren;Laurie Wiest;Anthony E. Pegg
亚精胺/精胺n1 -乙酰转移酶（Spd/Spm乙酰转移酶）是多胺分解代谢的限速酶。该酶可被多种刺激物高度诱导，包括天然多胺及其结构类似物。为了研究控制该酶的潜在机制，我们从多胺类似物处理的大细胞肺癌NCI H157的mRNA随机引物cDNA文库中分离出一个编码人类Spd/Spm乙酰转移酶的cDNA。利用从纯化蛋白中提取的7个氨基酸多肽序列，用32倍退化的20个碱基低聚物探针鉴定了972个碱基对的cDNA。该cDNA具有513个碱基的开放阅读框，编码171个氨基酸的蛋白质，预计分子量为20,023。体外翻译研究证明该cDNA的蛋白产物是一种具有生物活性的酶。该cDNA在人细胞中识别1.5千碱基的转录物，该转录物在人大细胞肺癌NCI H157细胞系中经多胺类似物处理后被高度诱导。NCI H157细胞对治疗的反应中Spd/Spm乙酰转移酶mRNA的异常高表达似乎不是Spd/Spm乙酰转移酶基因扩增的结果。
Spermidine/spermine N1-acetyltransferase (Spd/Spm acetyltransferase) is the rate-limiting enzyme in the catabolism of polyamines. This enzyme is highly inducible by several stimuli, including the natural polyamines and their structural analogues. To investigate the underlying mechanism responsible for the control of this enzyme a cDNA which codes for an active human Spd/Spm acetyltransferase has been isolated from a random primed cDNA library constructed from mRNA of a polyamine analogue treated large cell lung carcinoma line, NCI H157. The 972-base pair cDNA was identified using a 32-fold degenerate, 20-base oligomer probe to a 7-amino acid polypeptide sequence derived from the purified protein. The cDNA has a 513-base open reading frame that codes for a protein of 171 amino acids with a predicted molecular weight of 20,023. In vitro translation studies demonstrated the protein product of this cDNA to be a biologically active enzyme. The cDNA recognizes a 1.5-kilobase transcript in human cells which is highly induced in the human large cell lung carcinoma NCI H157 line following treatment with the polyamine analogue. The unusually high expression of Spd/Spm acetyltransferase mRNA by the NCI H157 cells in response to treatment does not appear to be a result of an amplification of the Spd/Spm acetyltransferase gene.