Margulies L, Wallace S S
Department of Biology, Mercy College, Dobbs Ferry, NY 10522.
Cell Biol Toxicol. 1984 Oct;1(1):127-43. doi: 10.1007/BF00125570.
An endonuclease activity that acts on alkali-labile lesions in x-irradiated PM2 DNA and recognizes apurinic lesions in heat/acid treated DNA has been partially purified from Drosophila melanogaster embryos and its specific activity monitored throughout early development. The enzyme activity also showed a low level of activity on UV-irradiated DNA. The saturation kinetics observed with both x-irradiated and apurinic PM2 DNA substrates were similar. The endonuclease activity exhibited a broad pH optimum between pH 6 and 8.5 and was almost completely inhibited by 100 mM NaCl, 0.1 mM EDTA, 2 mM CaCl2 and 10 mM NEM. The reaction was not completely dependent on the presence of Mg++cation, but optimum activity was obtained at a concentration of 0.1 mM; concentrations greater than 1 mM Mg++ were inhibitory. The specific activity of the apurinic endonuclease, partially purified from several stages of embryonic and early larval development, remained the same. Unfertilized eggs exhibited a reduced level of this presumptive repair activity.
一种作用于经X射线照射的PM2 DNA中对碱不稳定损伤的核酸内切酶活性,以及识别经热/酸处理的DNA中的脱嘌呤损伤的核酸内切酶活性,已从黑腹果蝇胚胎中部分纯化,并在整个早期发育过程中监测其比活性。该酶活性在紫外线照射的DNA上也表现出低水平的活性。用经X射线照射的和脱嘌呤的PM2 DNA底物观察到的饱和动力学相似。该核酸内切酶活性在pH 6至8.5之间表现出较宽的最适pH值,并且几乎完全被100 mM NaCl、0.1 mM EDTA、2 mM CaCl2和10 mM NEM抑制。该反应并不完全依赖于Mg++阳离子的存在,但在0.1 mM的浓度下可获得最佳活性;大于1 mM Mg++的浓度具有抑制作用。从胚胎和幼虫早期发育的几个阶段部分纯化的脱嘌呤核酸内切酶的比活性保持不变。未受精卵表现出这种假定修复活性的降低水平。