Cytoplasmic Dynein as a Facilitator of Nuclear Envelope Breakdown (original) (raw)
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Journal of Cell Biology, 1997
The mechanisms of localization and retention of membrane proteins in the inner nuclear membrane and the fate of this membrane system during mitosis were studied in living cells using the inner nuclear membrane protein, lamin B receptor, fused to green fluorescent protein (LBR-GFP). Photobleaching techniques revealed the majority of LBR-GFP to be completely immobilized in the nuclear envelope (NE) of interphase cells, suggesting a tight binding to heterochromatin and/or lamins. A subpopulation of LBR-GFP within ER membranes, by contrast, was entirely mobile and diffused rapidly and freely ( D ϭ 0.41 Ϯ 0.1 m 2 /s). High resolution confocal time-lapse imaging in mitotic cells revealed LBR-GFP redistributing into the interconnected ER membrane system in prometaphase, exhibiting the same high mobility and diffusion constant as observed in interphase ER membranes. LBR-GFP rapidly diffused across the cell within the membrane network defined by the ER, suggesting the integrity of the ER was maintained in mitosis, with little or no fragmentation and vesiculation. At the end of mitosis, nuclear membrane reformation coincided with immobilization of LBR-GFP in ER elements at contact sites with chromatin. LBR-GFP-containing ER membranes then wrapped around chromatin over the course of 2-3 min, quickly and efficiently compartmentalizing nuclear material. Expansion of the NE followed over the course of 30-80 min. Thus, selective changes in lateral mobility of LBR-GFP within the ER/NE membrane system form the basis for its localization to the inner nuclear membrane during interphase. Such changes, rather than vesiculation mechanisms, also underlie the redistribution of this molecule during NE disassembly and reformation in mitosis.
Genes to Cells, 2019
The nuclear envelope (NE) is a double-membrane structure composed of inner and outer nuclear membranes. The outer membrane is continuous with the endoplasmic reticulum (ER). The inner nuclear membrane contains various NEspecific integral membrane proteins that can interact with the nuclear lamina, chromatin and other NE proteins. The outer nuclear membrane shares proteins with those on the ER membrane. The nuclear pore complex (NPC) is a large protein complex found in the merged regions of the inner and outer nuclear membranes (reviewed in De Magistris & Antonin, 2018; Goldberg & Allen, 1995). Additionally, the nuclear lamina structure is present beneath the nuclear membrane in metazoan including humans (reviewed in de Leeuw,
Nuclear Membrane Disassembly and Rupture
Journal of Molecular Biology, 2007
The nuclear envelope consists of two membranes traversed by nuclear pore complexes. The outer membrane is continuous with the endoplasmic reticulum. At mitosis nuclear pore complexes are dismantled and membranes disperse. The mechanism of dispersal is controversial: one view is that membranes feed into the endoplasmic reticulum, another is that they vesiculate. Using Xenopus egg extracts, nuclei have been assembled and then induced to breakdown by addition of metaphase extract. Field emission scanning electron microscopy was used to study disassembly. Strikingly, endoplasmic reticulum-like membrane tubules form from the nuclear surface after the addition of metaphase extracts, but vesicles were also observed. Microtubule inhibitors slowed but did not prevent membrane removal, whereas Brefeldin A, which inhibits vesicle formation, stops membrane disassembly, suggesting that vesiculation is necessary. Structures that looked like coated buds were observed and buds were labelled for β-COP. We show that nuclear pore complexes are dismantled and the pore closed prior to membrane rupturing, suggesting that rupturing is an active process rather than a result of enlargement of nuclear pores.
Sequential recruitment of NPC proteins to the nuclear periphery at the end of mitosis
Journal of Cell Science, 1999
Nuclear pore complexes (NPCs) are extremely elaborate structures that mediate the bidirectional movement of macromolecules between the nucleus and cytoplasm. With a mass of about 125 MDa, NPCs are thought to be composed of 50 or more distinct protein subunits, each present in multiple copies. During mitosis in higher cells the nuclear envelope is disassembled and its components, including NPC subunits, are dispersed throughout the mitotic cytoplasm. At the end of mitosis, all of these components are reutilized. Using both conventional and digital confocal immunofluorescence microscopy we have been able to define a time course of post-mitotic assembly for a group of NPC components (CAN/Nup214, Nup153, POM121, p62 and Tpr) relative to the integral nuclear membrane protein LAP2 and the NPC membrane glycoprotein gp210. Nup153, a component of the nuclear basket, associates with chromatin towards the end of anaphase, in parallel with the inner nuclear membrane protein, LAP2. However, immu...
Regulation and coordination of nuclear envelope and nuclear pore complex assembly
Nucleus (Austin, Tex.)
In metazoans with "open" mitosis, cells undergo structural changes involving the complete disassembly of the nuclear envelope (NE). In post-mitosis, the dividing cell faces the difficulty to reassemble NE structures in a highly regulated fashion around separated chromosomes. The de novo formation of nuclear pore complexes (NPCs), which are gateways between the cytoplasm and nucleoplasm across the nuclear membrane, is an archetype of macromolecular assembly and is therefore of special interest. The reformation of a functional NE further involves the reassembly and organization of other NE components, the nuclear membrane and NE proteins, around chromosomes in late mitosis. Here, we discuss the function of NE components, such as lamins and INM proteins, in NE reformation and highlight recent results on coordination of NPC and NE assembly.
Nuclear envelope insertion of spindle pole bodies and nuclear pore complexes
Nucleus, 2012
T he defining feature of eukaryotic cells is the double lipid bilayer of the nuclear envelope (NE) that serves as a physical barrier separating the genome from the cytosol. Nuclear pore complexes (NPCs) are embedded in the NE to facilitate transport of proteins and other macromolecules into and out of the nucleus. In fungi and early embryos where the NE does not completely breakdown during mitosis, microtubule-organizing centers such as the spindle pole body (SPB) must also be inserted into the NE to facilitate organization of the mitotic spindle. Several recent papers have shed light on the mechanism by which SPB complexes are inserted into the NE. An unexpected link between the SPB and NPCs suggests that assembly of these NE complexes is tightly coordinated. We review the findings of these reports in light of our current knowledge of SPB, NPC and NE structure, assembly and function.