Cohrs R J, Goswami B B, Sharma O K
Department of Molecular Biology, AMC Cancer Research Center, Denver, Colorado 80214.
Biochemistry. 1988 May 3;27(9):3246-52. doi: 10.1021/bi00409a019.
Rapid withdrawal of estrogen from immature chicks, previously stimulated with the hormone, results in the inhibition of transcription of mRNAs of egg white proteins, rapid degradation of existing estrogen-induced mRNAs of egg white proteins, and decline in ribosomes and weight of the oviduct. On rapid withdrawal of estrogen, ovalbumin mRNA decreased to 65% after 3 h and was not detected after 24 h. In contrast to ovalbumin mRNA, cellular RNA content remained unchanged at 3 h and subsequently decreased to 51% of the stimulated value by 48 h. To study the mechanism of rapid degradation of RNA during estrogen withdrawal, the role of 2-5A- [px(A2'p)nA; x = 2 or 3, n greater than or equal to 2] dependent RNase was investigated. The effect of 2-5A-dependent RNase on the stability of RNA in vitro was determined by incubating oviduct polysomes with 2-5A-dependent RNase and exogenous 2-5A. Ovalbumin mRNA was degraded more rapidly than beta-actin mRNA and rRNA, and the kinetics of RNA degradation were very similar to those observed in vivo. Levels of 2-5A in the chick oviduct increased shortly after estrogen withdrawal. Analysis of the oviduct RNA revealed that a distinct 18S rRNA derived fragment, 450 nucleotides in length, increased at 6 h after withdrawal and at subsequent time points when significant degradation of total cellular RNA was occurring. The 18S rRNA derived degradation product observed in vivo from the chick oviduct had the same mobility in denaturing agarose gels as the 18S rRNA cleavage product liberated on incubation of isolated oviduct ribosomes with purified 2-5A-dependent RNase and exogenous 2-5A.(ABSTRACT TRUNCATED AT 250 WORDS)
对于先前用雌激素刺激过的未成熟雏鸡,迅速撤去雌激素会导致蛋清蛋白mRNA转录受到抑制、现有的雌激素诱导的蛋清蛋白mRNA迅速降解,以及输卵管核糖体数量和重量下降。迅速撤去雌激素后,卵清蛋白mRNA在3小时后降至65%,24小时后未检测到。与卵清蛋白mRNA不同,细胞RNA含量在3小时时保持不变,随后到48小时降至刺激值的51%。为研究撤去雌激素期间RNA快速降解的机制,研究了2-5A- [px(A2'p)nA;x = 2或3,n大于或等于2]依赖性核糖核酸酶的作用。通过将输卵管多核糖体与2-5A依赖性核糖核酸酶和外源性2-5A一起孵育,测定2-5A依赖性核糖核酸酶对体外RNA稳定性的影响。卵清蛋白mRNA比β-肌动蛋白mRNA和rRNA降解得更快,RNA降解动力学与体内观察到的非常相似。撤去雌激素后不久,雏鸡输卵管中2-5A的水平升高。对输卵管RNA的分析显示,一个长度为450个核苷酸的独特18S rRNA衍生片段在撤去雌激素后6小时以及随后细胞总RNA发生显著降解的时间点增加。在雏鸡输卵管体内观察到的18S rRNA衍生降解产物在变性琼脂糖凝胶中的迁移率与分离的输卵管核糖体与纯化的2-5A依赖性核糖核酸酶和外源性2-5A一起孵育时释放的18S rRNA切割产物相同。(摘要截短至250字)