PYRIDINE NUCLEOTIDES--TURNOVER AND REGULATION

本研究旨在了解吡啶核苷酸周转和 监管。下一个资助期的主要工作将是理解 NAD在真核生物中的核代谢，特别是作为底物 用于聚(ADP-核糖基化)。DNA的生物化学特性 刺激和结合部位，以及多聚ADP-核糖连接部位 有计划的。此外，还编制了标记纯化的微量注射程序 酶进入活的哺乳动物细胞的培养将被进行。 生物化学和遗传学相结合的方法也将在 鼠伤寒沙门氏菌。所有剩余的未定义基因都参与其中 在吡啶中的核苷酸将被鉴定并在这些基因座上突变 获得并表征了。吡啶核苷酸基因的突变体 循环将被用来开发一种简便的方法来测试DNA连接 活着。吡啶核苷酸生物合成途径的调控 将对营业额进行调查；初步方法将是建造 在所有已知的基因中都有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.