Bast R E, Garfield S A, Gehrke L, Ilan J
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3133-7. doi: 10.1073/pnas.74.8.3133.
To elucidate the mechanisms by which protein synthesis is affected by estradiol, we characterized cockerel liver polysomal profiles during hormone induction and withdrawal. We describe a method for isolating intact polysomes which results in preparations that are stable even after storage in solution at 10 degrees for 16 hr. In addition, our procedure eliminates the necessity for starving animals prior to experiments. Recovery of radioactive polysomes indicated that yield is about 90% and that our polysomal preparations appear to represent polysome distribution in vivo. Using this approach we show that estradiol injection stimulates ribosome content 6-fold and that formation of polysomes is coincident with the induction of vitellogenin synthesis. We also demonstrate that the size and number of polysomes increase and decrease in a coordinated fashion with the rate of vitellogenin synthesis. The kinetics of ribosome synthesis and the fact that at least 80% of the newly synthesized ribosomes are directly recruited into polysomes indicate that ribosomes might be limiting the rate of protein synthesis during the stimulatory phase of the hormone cycle.
为阐明雌二醇影响蛋白质合成的机制,我们对激素诱导和撤除过程中的公鸡肝脏多核糖体图谱进行了表征。我们描述了一种分离完整多核糖体的方法,该方法所得到的制剂即使在10摄氏度的溶液中储存16小时后仍保持稳定。此外,我们的方法无需在实验前使动物饥饿。放射性多核糖体的回收率表明,产量约为90%,并且我们的多核糖体制剂似乎代表了体内的多核糖体分布。使用这种方法,我们发现注射雌二醇可使核糖体含量增加6倍,并且多核糖体的形成与卵黄蛋白原合成的诱导同时发生。我们还证明,多核糖体的大小和数量随着卵黄蛋白原合成速率以协调的方式增加和减少。核糖体合成的动力学以及至少80%新合成的核糖体直接被招募到多核糖体中的事实表明,在激素周期的刺激阶段,核糖体可能限制了蛋白质合成的速率。