Müller W E, Rohde H J, Zahn R K
Biochimie. 1976;58(5):543-50. doi: 10.1016/s0300-9084(76)80224-6.
The biological model of the selective induction of RNA synthesis in oviducts of estrogen stimulated immature quails by progesterone has been used to clarify whether poly (AD-Rib) is involved in DNA transcription. The chromatin-bound as well as the soluble poly (ADP-Rib) polymerase has been isolated from oviducts and the optimal reaction conditions have been determined. The activities, as measured by the incorporation rates of NAD+ into poly (ADP-Rib), of both, chromatin-bound "endogenous" polymerase (in the absense of "exogenous" DNA and histones) and soluble enzyme (native DNA-lysine-rich histone ratio: 4.3) from progesterone treated quail oviducts, have been determined to be only 30 per cent and 46 per cent respectively, as compared with the activities of the enzymes from the controls. This decrease in incorporation rates is apparently not due to an increased poly (ADP-Rib) degrading enzyme activity. Poly (ADP-Rib) synthesis in vivo was determined by incorporation studies with the precursor (14C) ribose. 12 h after intraperitoneal administration, 0.014 per cent of the total radioactivity was recovered in the oviduct histone fraction, 0.011 per cent in the oviduct nonhistone fraction and 0.009 per cent in the oviduct "HCIextract" containing the histone subfractions f1, f2 and f3. Among these histone subfractions f1 is ADP-ribosylated to the largest extent. ADP-ribosylation of f1 is less extensive in progesterone-stimulated oviducts (65 per cent) than in the controls (100 per cent). The present results suggest that in course of the selective, progesterone-induced DNA transcription the poly (ADP-Rib synthesis might drop.
通过孕酮对雌激素刺激的未成熟鹌鹑输卵管中RNA合成进行选择性诱导的生物学模型,已被用于阐明聚(ADP - 核糖)是否参与DNA转录。已从输卵管中分离出与染色质结合的以及可溶性的聚(ADP - 核糖)聚合酶,并确定了最佳反应条件。通过NAD⁺掺入聚(ADP - 核糖)的速率来衡量,与对照组的酶活性相比,孕酮处理的鹌鹑输卵管中与染色质结合的“内源性”聚合酶(在不存在“外源性”DNA和组蛋白的情况下)和可溶性酶(天然DNA与富含赖氨酸的组蛋白比例为4.3)的活性分别仅为30%和46%。掺入率的这种降低显然不是由于聚(ADP - 核糖)降解酶活性的增加。体内聚(ADP - 核糖)的合成通过用前体(¹⁴C)核糖进行掺入研究来确定。腹腔注射后12小时,在输卵管组蛋白部分中回收了总放射性的0.014%,在输卵管非组蛋白部分中回收了0.011%,在含有组蛋白亚组分f1、f2和f3的输卵管“HCl提取物”中回收了0.009%。在这些组蛋白亚组分中,f1的ADP - 核糖基化程度最大。在孕酮刺激的输卵管中,f1的ADP - 核糖基化程度(65%)低于对照组(100%)。目前的结果表明,在孕酮诱导的选择性DNA转录过程中,聚(ADP - 核糖)的合成可能会下降。