Shirasawa N, Matsuno T
Biochim Biophys Acta. 1979 Apr 26;562(2):271-80. doi: 10.1016/0005-2787(79)90172-2.
The suppression mechanism of glutamine synthetase [EC 6.3.1.2] induction mediated by chick interferon preparation was investigated in embryonic chick neural retina cell cultures. A translational assay was used to measure the level of mRNA coding for the enzyme. RNA extracted from the retinal polysomes was chromatographed on oligo(dT)-cellulose and translated in a cell-free protein synthesizing system derived from wheat germ. The newly synthesized enzyme was isolated by immunoprecipitation with anti-enzyme gamma-globulin and identified using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Neither polysomal profiles nor total protein synthesis directed by the RNA preparations was affected by interferon treatment. Comparing the amount of glutamine synthetase synthesized in vitro by equal amounts of mRNA fraction from retinas treated with or without interferon indicated that the suppression was associated with reduced level of the enzyme mRNA on polysomes.
在鸡胚神经视网膜细胞培养物中研究了鸡干扰素制剂介导的谷氨酰胺合成酶[EC 6.3.1.2]诱导的抑制机制。采用翻译分析法来测量编码该酶的mRNA水平。从视网膜多核糖体中提取的RNA在寡聚(dT)-纤维素上进行层析,并在源自小麦胚芽的无细胞蛋白质合成系统中进行翻译。通过用抗酶γ-球蛋白进行免疫沉淀分离新合成的酶,并使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行鉴定。多核糖体图谱以及RNA制剂指导的总蛋白质合成均不受干扰素处理的影响。比较用或不用干扰素处理的视网膜等量mRNA组分在体外合成的谷氨酰胺合成酶的量,表明这种抑制与多核糖体上该酶mRNA水平的降低有关。