PYRIDINE NUCLEOTIDE: TURNOVER AND REGULATION

本研究旨在了解吡啶核苷酸周转和 调控 下一个资助期的一项主要工作将是了解 真核生物中NAD的核代谢，特别是作为底物 对于聚（ADP-核糖基化）。 DNA的一种生化表征 刺激和结合位点，以及聚ADP-核糖附着位点， 计划好了 此外，还建立了标记纯化的 将酶导入培养的活哺乳动物细胞中。 生物化学和遗传学相结合的方法也将继续在 鼠伤寒沙门氏菌 所有剩下的未知基因 在吡啶中，将鉴定核苷酸和这些基因座突变体 获得并表征。 吡啶核苷酸基因突变 循环将被用来开发一种方便的方法来测定DNA连接， vivo. 吡啶核苷酸生物合成途径的调节， 营业额将进行调查;最初的方法将是建立 所有已知基因中的Mu-lac融合。

This research aims at understanding pyridine nucleotide turnover and regulation. A major effort in the next grant period will be to understand the nuclear metabolism of NAD in eukaryotes, specifically as a substrate for poly (ADP-ribosylation). A biochemical characterization of DNA stimulatory and binding sites, and poly ADP-ribose attachment sites is planned. In addition, a program of microinjection of labeled purified enzymes into living mammalian cells in culture will be carried out. A combined biochemical and genetic approach will also be continued in the bacterium Salmonella typhimurium. All remaining undefined genes involved in pyridine nucleotide will be identified and mutants at these loci obtained and characterized. Mutants in genes of the pyridine nucleotide cycle will be used to develop a convenient method to assay DNA ligation in vivo. The regulation of pyridine nucleotide pathways for biosynthesis and turnover will be investigated; the initial approach will be to construct Mu-lac fusions in all known genes.