RUI: The Mechanism of mRNP Activation at Fertilization

这项建议的主要目的是阐明控制海胆卵受精时蛋白质合成激活的机制。蛋白质合成的增加是通过激活储存的母体信使核糖核蛋白(MRNPs)和增加翻译机制的活性来调节的。格兰杰博士建议对控制储存的母体mRNPs抑制和激活的因素进行详细研究。通过比较非活性鸡蛋Poly(A)+mRNPs和活性受精Poly(A)+mRNPs的蛋白质模式，将识别参与鸡蛋mRNPs翻译抑制的蛋白质。此外，将从体外激活的mRNPs中分离出潜在的掩蔽因子候选者。当卵子mRNP在受精时被激活时，针对这些mRNP掩蔽蛋白的抗体将被用来监测mRNP蛋白的变化。细胞内钙和pH的变化会引起细胞的激活和蛋白质合成速度的加快。格兰杰博士将改变未受精卵的钙和pH水平，并测量这些变化对鸡蛋mRNPs翻译活性的影响。使用针对mRNP掩蔽因子的抗体，他将确定这些治疗修改或释放了哪些蛋白质。未受精卵是一种代谢静止、不分裂的细胞。受精导致一系列途径的激活，包括那些导致蛋白质合成速度加快的途径。格兰杰博士建议以海胆为模型系统，研究受精后蛋白质合成激活所涉及的生化过程。

The broad objective of this proposal is to elucidate the mechanisms that control the activation of protein synthesis at fertilization in sea urchin eggs. The increase in protein synthesis is mediated by both the activation of stored maternal messenger ribonucleoprotein (mRNPs) and an increase in the activity of the translational machinery. Dr. Grainger proposes to do a detailed study of the factors that control the repression and activation of stored maternal mRNPs. Proteins that are involved in the translational repression of egg mRNPs will be identified by comparing the protein pattern of inactive egg poly(A)+mRNPs and active fertilized poly(A)+mRNPs. In addition, potential candidates for masking factors will be isolated from in vitro activated mRNPs. Antibodies against these mRNP masking proteins will be used to monitor changes in mRNP proteins as egg mRNPs become activated at fertilization. Changes in intracellular calcium and pH trigger ugg activation and the increase in the rate of protein synthesis. Dr. Grainger will alter the calcium and pH levels of unfertilized eggs and measure the effect of these changes on the translational activity of egg mRNPs. Using antibodies against the mRNP masking factors, he will determine which proteins are modified or released by these treatments. %%% The unfertilized egg is a metabolically quiescent, non- dividing cell. Fertilization results in the activation of a series of pathways, including those leading to an increase in the rate of protein synthesis. Dr. Grainger proposes to study the biochemical processes involved in this activation of protein synthesis upon fertilization using the sea urchin as a model system.