Temporal differences in the appearance of NEP-B78 and an LBR-like protein during Xenopus nuclear envelope reassembly reflect the ordered recruitment of functionally discrete vesicle types
Temporal differences in the appearance of NEP-B78 and an LBR-like protein during Xenopus nuclear envelope reassembly reflect the ordered recruitment of functionally discrete vesicle types
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DOI：
10.1083/jcb.144.2.225
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发表时间：
1999-01-25
影响因子：
7.8
通讯作者：
Hutchison, CJ
中科院分区：
文献类型：
--
作者：
Drummond, S;Ferrigno, P;Hutchison, CJ
在这项工作中，我们已经使用了新的单克隆抗体对两种蛋白质的内质网和外核膜，称为NEP-B78和p65，除了对内核膜蛋白LBR（核纤层蛋白B受体）的多克隆抗体，研究的顺序和动力学的NE重组在非洲爪蟾无细胞系统。使用这些试剂，我们证明了在无细胞系统和XLK-2细胞中，其同源膜蛋白募集到去致密染色质表面的时间差异。我们明确地表明，在无细胞系统中，两种功能和生化不同的囊泡类型是必要的NE组装。我们发现，不同的囊泡招聘染色质的过程是一个有序的，NEP-B78定义了一个囊泡人口参与最早的事件在这个系统中的重组。最后，我们提出的证据表明，NEP-B78可能需要这些囊泡的目标，在这个系统中的去致密染色质的表面。这些结果对于理解有丝分裂过程中核膜拆卸和重新组装的机制以及开发识别控制这些过程的新分子的系统具有重要意义。
In this work, we have used novel mAbs against two proteins of the endoplasmic reticulum and outer nuclear membrane, termed NEP-B78 and p65, in addition to a polyclonal antibody against the inner nuclear membrane protein LBR (lamin B receptor), to study the order and dynamics of NE reassembly in the Xenopus cell-free system. Using these reagents, we demonstrate differences in the timing of recruitment of their cognate membrane proteins to the surface of decondensing chromatin in both the cell-free system and XLK-2 cells. We show unequivocally that, in the cell-free system, two functionally and biochemically distinct vesicle types are necessary for NE assembly. We find that the process of distinct vesicle recruitment to chromatin is an ordered one and that NEP-B78 defines a vesicle population involved in the earliest events of reassembly in this system. Finally, we present evidence that NEP-B78 may be required for the targeting of these vesicles to the surface of decondensing chromatin in this system. The results have important implications for the understanding of the mechanisms of nuclear envelope disassembly and reassembly during mitosis and for the development of systems to identify novel molecules that control these processes.