Chambers S A, Vaughn J P, Shaw B R
Biochemistry. 1983 Nov 22;22(24):5626-31. doi: 10.1021/bi00293a026.
Prior to fertilization, sperm possess one of the longest nucleosome repeat lengths yet determined [approximately 250 base pairs (bp) for the sea urchin Strongylocentrotus purpuratus]. We show here that the two-cell embryo has an average repeat size of 189 +/- 2 bp as probed by micrococcal nuclease; this is the shortest average nucleosomal subunit reported for S. purpuratus. By the eight-cell stage, the average nucleosome repeat increases to 201 +/- 2 bp, and it subsequently increases further during development. These results indicate that a dramatic rearrangement of chromatin occurs upon fertilization and that this chromatin remodeling continues through early development. When two-cell embryos are labeled for 30 min with [3H]thymidine and digested briefly, they exhibit nuclease-hypersensitive fragments averaging 308 bp in size, which are consistent with the size of protected DNA units in replication intermediate complexes at blastula stage (as described by Levy and Jacob [Levy, A., & Jacob, K. M. (1978) Cell (Cambridge, Mass.) 14, 259]). Our results are consistent with two general propositions: (1) long repeat lengths are found in highly differentiated cells, and (2) short repeat lengths are characteristic of cells more active in cell division. Our data would also imply that a rapid increase in the DNA complement, e.g., in the transition from haploid to diploid state following fertilization, is accompanied by a shortening of the average size of DNA in a nucleosome after replication.
在受精之前,精子拥有迄今所测定的最长核小体重复长度之一(海胆紫球海胆约为250个碱基对)。我们在此表明,经微球菌核酸酶检测,两细胞胚胎的平均重复大小为189±2个碱基对;这是报道的紫球海胆最短的平均核小体亚基。到八细胞阶段,平均核小体重复长度增加到201±2个碱基对,并且在随后的发育过程中进一步增加。这些结果表明,受精时染色质发生了剧烈重排,并且这种染色质重塑在早期发育过程中持续存在。当用[³H]胸腺嘧啶核苷标记两细胞胚胎30分钟并进行短暂消化时,它们会呈现出平均大小为308个碱基对的核酸酶超敏片段,这与囊胚期复制中间复合物中受保护的DNA单元大小一致(如Levy和Jacob所述[Levy, A., & Jacob, K. M. (1978) Cell (Cambridge, Mass.) 14, 259])。我们的结果与两个一般性观点一致:(1)在高度分化的细胞中发现长重复长度,(2)短重复长度是细胞分裂更活跃的细胞的特征。我们的数据还意味着,DNA互补物的快速增加,例如在受精后从单倍体状态向二倍体状态的转变过程中,伴随着复制后核小体中DNA平均大小的缩短。