Yoshihara K, Itaya A, Tanaka Y, Ohashi Y, Ito K, Teraoka H, Tsukada K, Matsukage A, Kamiya T
Biochem Biophys Res Commun. 1985 Apr 16;128(1):61-7. doi: 10.1016/0006-291x(85)91644-4.
Incubation of DNA polymerase alpha, DNA polymerase beta, terminal deoxynucleotidyl transferase, or DNA ligase II in a reconstituted poly(ADP-ribosyl)ating enzyme system markedly suppressed the activity of these enzymes. Components required for poly(ADP-ribose) synthesis including poly(ADP-ribose) polymerase, NAD+, DNA, and Mg2+ were all essential for the observed suppression. Purified poly(ADP-ribose) itself, however, was slightly inhibitory to all of these enzymes. Furthermore, the suppressed activities of DNA polymerase alpha, DNA polymerase beta, and terminal deoxynucleotidyl transferase were largely restored (3 to 4-fold stimulation was observed) by a mild alkaline treatment, a procedure known to hydrolyze alkaline-labile ester linkage between poly(ADP-ribose) and an acceptor protein. All of these results strongly suggest that the four nuclear enzymes were inhibited as a result of poly(ADP-ribosyl)ation of either the enzyme molecule itself or some regulatory proteins of these enzymes.
在重组的聚（ADP-核糖基）化酶系统中孵育DNA聚合酶α、DNA聚合酶β、末端脱氧核苷酸转移酶或DNA连接酶II，可显著抑制这些酶的活性。聚（ADP-核糖）合成所需的成分，包括聚（ADP-核糖）聚合酶、NAD⁺、DNA和Mg²⁺，对于观察到的抑制作用都是必不可少的。然而，纯化的聚（ADP-核糖）本身对所有这些酶有轻微的抑制作用。此外，通过温和的碱性处理（一种已知可水解聚（ADP-核糖）与受体蛋白之间碱不稳定酯键的方法），DNA聚合酶α、DNA聚合酶β和末端脱氧核苷酸转移酶被抑制的活性在很大程度上得以恢复（观察到3至4倍的刺激）。所有这些结果都强烈表明，这四种核酶是由于酶分子本身或这些酶的一些调节蛋白的聚（ADP-核糖基）化而被抑制的。
