Schröder J, Kreuzaler F, Schäfer E, Hahlbrock K
J Biol Chem. 1979 Jan 10;254(1):57-65.
Irradiation of previously dark-grown cell suspension cultures from parsley (Petroselinum hortense Hoffm.) with ultraviolet light caused large, concomitant increases in mRNA activities for two characteristic enzymes of phenylpropanoid metabolism, phenylalanine ammonia-lyase, and flavanone synthase. The rates of enzyme synthesis both in vitro in a reticulocyte lysate and in vivo were quantitated by direct immunoprecipitation of the labeled enzyme subunits. Following a period of about 2 h, during which possible changes were below the limits of detectability, the two mRNA activities increased rapidly in irradiated cells for serveral hours. Phenylalanine ammonia-lyase mRNA reached a peak in activity a few hours earlier than flavanone synthase mRNA. The apparent half-lives of the enzyme activities were about 7 to 10 h for phenylalanine ammonia-lyase and 5 to 7 h for flavanone synthase. These data were used to calculate the expected, light-induced changes in enzyme activity from the measured changes in mRNA activity. The results for both enzymes were in agreement with experimental data, indicating that the light-induced changes in enzyme activity can be explained by changes in the respective mRNA activity. Some data concerning changes in the degree of polyadenylation of the mRNAs and the inhibitory effects of 7-methylguanosine 5'-phosphate are presented.
用紫外线照射之前在黑暗中培养的欧芹(Petroselinum hortense Hoffm.)细胞悬浮培养物,导致苯丙烷类代谢的两种特征酶——苯丙氨酸解氨酶和黄烷酮合酶的mRNA活性大幅增加,且二者同时出现这种情况。通过对标记的酶亚基进行直接免疫沉淀,对网织红细胞裂解物中的体外酶合成速率和体内酶合成速率进行了定量。在大约2小时的时间段内,可能的变化低于可检测限度,之后在照射的细胞中,这两种mRNA活性迅速增加了几个小时。苯丙氨酸解氨酶mRNA的活性峰值比黄烷酮合酶mRNA早几个小时出现。苯丙氨酸解氨酶的酶活性表观半衰期约为7至10小时,黄烷酮合酶的酶活性表观半衰期约为5至7小时。利用这些数据,根据测得的mRNA活性变化计算出预期的光诱导酶活性变化。两种酶的结果均与实验数据一致,表明光诱导的酶活性变化可以用各自mRNA活性的变化来解释。还给出了一些关于mRNA聚腺苷酸化程度变化以及5'-磷酸-7-甲基鸟苷抑制作用的数据。