Imschenetzky M, Montecino M, Puchi M
Universidad de Concepción, Chile.
J Cell Biochem. 1991 Jul;46(3):234-41. doi: 10.1002/jcb.240460306.
The patterns of poly(ADP-ribosylation) in vivo of CS (cleavage stage) histone variants were compared in sea urchin zygotes at the entrance and the exit of S1 and S2 in the initial developmental cell cycles. This post-translational modification was detected by Western immunoblots with rabbit sera anti-poly(ADP-ribose) that was principally reactive against ADP-ribose polymers and slightly against ADP-ribose oligomers. The effect of 3 aminobenzamide (3-ABA), an inhibitor of the poly(ADP-ribose) synthetase, on S phase progression was determined in vivo by measuring the incorporation of 3H thymidine into DNA. The results obtained indicate that the CS histone variants are poly(ADP-ribosylated) in a cell cycle dependent manner. A significantly positive reaction of several CS variants with sera anti-poly(ADP-ribose) was found at the entrance into S phase, which decreases after its completion. The incubation of zygotes in 3-ABA inhibited the poly(ADP-ribosylation) of CS variants and prevented both the progression of the first S phase and the first cleavage division. These observations suggest that the poly(ADP-ribosylation) of atypical CS histone variants is relevant for initiation of sea urchin development and is required for embryonic DNA replication.
在海胆受精卵最初发育细胞周期的S1和S2期进入和退出时,比较了CS(卵裂期)组蛋白变体在体内的多聚(ADP-核糖基化)模式。通过用主要与ADP-核糖聚合物反应且对ADP-核糖寡聚物反应较弱的抗多聚(ADP-核糖)兔血清进行Western免疫印迹检测这种翻译后修饰。通过测量3H胸苷掺入DNA来在体内确定多聚(ADP-核糖)合成酶抑制剂3-氨基苯甲酰胺(3-ABA)对S期进程的影响。所得结果表明CS组蛋白变体以细胞周期依赖性方式进行多聚(ADP-核糖基化)。在进入S期时发现几种CS变体与抗多聚(ADP-核糖)血清有明显阳性反应,在S期完成后这种反应减弱。受精卵在3-ABA中孵育会抑制CS变体的多聚(ADP-核糖基化),并阻止第一个S期进程和第一次卵裂。这些观察结果表明,非典型CS组蛋白变体的多聚(ADP-核糖基化)与海胆发育的起始相关,并且是胚胎DNA复制所必需的。