Regulation of Photosynthetic Electron Transport Genes in Chlamydomonas Chloroplasts

RNA稳定性的调控在基因表达中起着重要作用 表达，但所涉及的机制知之甚少。 该提案涉及分子和生物化学因素， 调节高等植物质体mRNA的差异周转， 细胞器生物发生 重点放在研究高度 保守的反向重复（IR）序列发现在3'端的 质体mRNA。 质体3'IR结合蛋白质并充当RNA 体外加工元件和稳定性决定因素。 我提议 研究IR中蛋白质：RNA的结合是否 参与质体RNA周转的调节。 这些研究 将需要3' IR序列的诱变，和 RNA结合蛋白这些蛋白质将用于功能 检测和提高抗体，这将用于调查 蛋白质浓度的发展变化，并最终在 分离编码它们的基因。 的体内结构 将通过分析核糖核蛋白来探测质体mRNA 颗粒（RNP），它们的半衰期将是 通过体内标记进行定量。 长期目标是 建立质体mRNA发育调控模型 稳定性，并确定核基因产物是否在 影响质体mRNA衰变的因素。*//

Regulation of RNA stability plays an important role in gene expression, but the mechanisms involved are poorly understood. This proposal addresses molecular and biochemical factors that regulate differential turnover of higher plant plastid mRNAs during organelle biogenesis. Emphasis is placed on the study of highly conserved inverted repeat (IR) sequences found at the 3' ends of plastid mRNAs. Plastid 3' IRs bind proteins, and act as RNA processing elements and stability determinants in vitro. I propose to investigate whether protein:RNA associations at the IR are involved in the regulation of plastid RNA turnover. These studies will entail mutagenesis of 3' IR sequences, and purification of RNA-binding proteins These proteins will be used in functional assays and to raise antibodies, which will be used to investigate developmental changes in protein concentrations, and eventually in isolation of the genes encoding them. The in vivo structure of plastid mRNAs will be probed by analyzing ribonucleoprotein particles (RNPs) from plastids, and their half lives will be quantified by in vivo labeling. The long-term objectives are to develop a model for the developmental control of plastid mRNA stability, and to determine if nuclear gene products are among the factors that influence plastid mRNA decay.***//