Nuclear reassembly in vitro is independent of nucleosome/chromatin assembly.
It was shown that nuclear reassembly was induced by small pieces of DNA fragments in cell-free extracts of Xenopus. In an attempt to learn the relationship between the nuclear reassembly and nucleosome/chromatin assembly, limited amounts of CM-Cellulose are used to eliminate the capacity of the egg extract S-150 to assemble chromatin. while the forming of nucleosomes is checked with DNA supercoiling by plasmid DNA pBR322 incubated in the extract, and further analysed by micrococcal nuclease digestion. This depleted extract is then used to induce nuclear reassembly around demembraned sperms with membrane vesicles. It is found that CM-Cellulose depletes histones H2A and H2B efficiently and blocks the assembly of nucleosomes, the demembraned sperms are yet reconstituted into nuclei in the treated S-150, although the chromatin in reassembled nuclei does not produce protected DNA fragments when digested with micrococcal nuclease. It suggests that in the cell-free system of Xenopus, DNA can be formed into nuclei without assembly of nucleosomes or chromatin.
Jiang
Z
Z
Peking University, Beijing, China.
Zhang
B
B
Zhai
Z
Z