NUCLEAR RECONSTITUTION INVITRO - STAGES OF ASSEMBLY AROUND PROTEIN-FREE DNA
NUCLEAR RECONSTITUTION INVITRO - STAGES OF ASSEMBLY AROUND PROTEIN-FREE DNA
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DOI：
10.1016/0092-8674(87)90424-7
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发表时间：
1987-01-30
期刊：
影响因子：
64.5
通讯作者：
NEWPORT, J
中科院分区：
文献类型：
--
作者：
NEWPORT, J
We have developed a cell-free system derived from Xenopus eggs that reconstitutes nuclear structure around an added protein-free substrate (bacteriophage lambda DNA). Assembled nuclei are morphologically indistinguishable from nomal eukaryotic nuclei: they are surrouned by a double membrane containing nuclear pores and are lined with a peripheral nuclear lamina. Nuclear assembly involves discrete intermediate steps, including nucleosome assembly, scaffold assembly, and nuclear membrane and lamina assembly, indicating that during reconstitution nuclear organization is assembled one level at a time. Topoisomerase II inhibitors block nuclear assembly. Lamin proteins and membrane vesicles bind to chromatin late in assembly, suggesting that these components do not interact with chromatin that is formed early in assembly. Reconstituted nuclei replicate their DNA: replication begins only after envelope formation has initiated, indicating that envelope attachment may be important for regulating replication.
