Temporal Differences in the Appearance of NEP-B78 and an LBR-like Protein during <i>Xenopus</i> Nuclear Envelope Reassembly Reflect the Ordered Recruitment of Functionally Discrete Vesicle Types

Drummond, Sheona P; Ferrigno, Paul Ko; Lyon, Carol E.; Murphy, Jane E.; Goldberg, Martin W.; Allen, Terence D; Smythe, Carl; Hutchison, Christopher J.

doi:10.1083/jcb.144.2.225

Cited by 66 publications

(65 citation statements)

References 42 publications

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“…Whether this is due to lateral diffusion within the membrane or to vesicle formation and fusion is an open question. In certain cases, extensively exemplified by Xenopus eggs, integral membrane proteins of the nuclear membrane are found in distinct vesicles (Vigers and Lohka, 1991;Buendia and Courvalin, 1997;Drummond et al, 1999). Indeed, recent reports suggest that there are multiple populations of specialized vesicles (Antonin et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Nuclear Envelope Breakdown Is Coordinated by Both Nup358/RanBP2 and Nup153, Two Nucleoporins with Zinc Finger Modules

Prunuske

Liu

Elgort

et al. 2006

MBoC

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When higher eukaryotic cells transition into mitosis, the nuclear envelope, nuclear pore complexes, and nuclear lamina are coordinately disassembled. The COPI coatomer complex, which plays a major role in membrane remodeling at the Golgi, has been implicated in the process of nuclear envelope breakdown and requires interactions at the nuclear pore complex for recruitment to this new site of action at mitosis. Nup153, a resident of the nuclear pore basket, was found to be involved in COPI recruitment, but the molecular nature of the interface between COPI and the nuclear pore has not been fully elucidated. To better understand what occurs at the nuclear pore at this juncture, we have probed the role of the nucleoporin Nup358/RanBP2. Nup358 contains a repetitive zinc finger domain with overall organization similar to a region within Nup153 that is critical to COPI association, yet inspection of these two zinc finger domains reveals features that also clearly distinguish them. Here, we found that the Nup358 zinc finger domain, but not a zinc finger domain from an unrelated protein, binds to COPI and dominantly inhibits progression of nuclear envelope breakdown in an assay that robustly recapitulates this process in vitro. Moreover, the Nup358 zinc finger domain interferes with COPI recruitment to the nuclear rim. Consistent with a role for this pore protein in coordinating nuclear envelope breakdown, Nup358-specific antibodies impair nuclear disassembly. Significantly, targeting either Nup153 or Nup358 for inhibition perturbs nuclear envelope breakdown, supporting a model in which these nucleoporins play nonredundant roles, perhaps contributing to COPI recruitment platforms on both the nuclear and cytoplasmic faces of the pore. We found that an individual zinc finger is the minimal interface for COPI association, although tandem zinc fingers are optimal. These results provide new information about the critical components of nuclear membrane remodeling and lay the foundation for a better understanding of how this process is regulated. INTRODUCTIONThe nuclear envelope creates a barrier that is critical to maintenance of the environment in the nucleus, specialized to support transcription and DNA replication. This double lipid membrane bilayer consists of an outer nuclear membrane, which is continuous with the endoplasmic reticulum (ER), and an inner nuclear membrane, which contains at least 78 different proteins, anchored via protein-protein interactions to the underlying nuclear lamina and chromatin . The nuclear envelope is perforated by nuclear pores, through which communication between the cytoplasm and nucleus occurs (Suntharalingam and Wente, 2003;Weis, 2003;Fahrenkrog et al., 2004;Pante, 2004). Despite the large size of the macromolecular nuclear pore complex (125 MDa in vertebrates), it is comprised of only ϳ30 different proteins, or nucleoporins (Nups), each present in multiple copies (Rout et al., 2000;Cronshaw et al., 2002). In metazoan cells, all these components-the lamina, nuclear pores, as well as the ...

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Section: Introductionmentioning

confidence: 99%

Nuclear Envelope Breakdown Is Coordinated by Both Nup358/RanBP2 and Nup153, Two Nucleoporins with Zinc Finger Modules

Prunuske

Liu

Elgort

et al. 2006

MBoC

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“…By biochemical methods, it was also shown that nuclear vesicles in mitosis were heterogeneous (Vigers and Lohka, 1992;Chaudhary and Courvalin, 1993;Lourim and Krohne, 1993;Buendia and Courvalin, 1997;Drummond et al, 1999). In the present paper, we prepared antibody against p60, a protein in LVs, to confirm the biochemical differences between LVs and HVs.…”

Section: Discussionmentioning

confidence: 67%

“…3B, lower). However, LBR-containing vesicles were incorporated into the nuclear envelope (Drummond et al, 1999). Thus, LBRcontaining HVs must bind to sperm-binding LVs to incorporate into the nuclear envelope before fusion events occur.…”

Section: Salt-treated Hvs Bind To Lvsmentioning

confidence: 99%

“…When sperm chromatin is incubated in the mixture of cytosol and purified membrane vesicles, an intact nuclear envelope is formed around the chromatin (Forbes et al, 1983;Lohka and Masui, 1983;Newport, 1987). According to analysis of membrane fractions from cell-free extracts, the population of nuclear vesicles required for nuclear envelope assembly might have biochemically and functionally heterogeneous compositions that are derived from either the inner, outer or pore nuclear membrane regions (Vigers and Lohka, 1992;Chaudhary and Courvalin, 1993;Lourim and Krohne, 1993;Gerace and Foisner, 1994;Buendia and Courvalin, 1997;Marshall and Wilson, 1997;Drummond et al, 1999). In this system, pretreatment of membrane vesicles with N-ethyl-maleimide (NEM) induces the inhibition of nuclear vesicle fusion, but the binding of nuclear vesicles to the chromatin occurs normally (Macaulay and Forbes, 1996).…”

Section: Introductionmentioning

confidence: 99%

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The interaction of two classes of nuclear vesicles is induced by the dissociation of soluble proteins from one class of vesicles

Oke

Inoue

2003

Biology of the Cell

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The regulatory mechanism of fusion steps in nuclear envelope assembly was studied in vitro using cell-free extracts of Xenopus eggs. The nuclear envelope is formed only around the chromatin at the end of mitosis. Two kinds of vesicles are required for nuclear envelope assembly (J. Cell Biol. 112 (1991) 545). Light vesicles (LVs) have the chromatin-binding activity, but cannot fuse solely. On the other hand, heavy vesicles (HVs) neither bind to the chromatin nor LVs, but are needed for the fusion event. Therefore, the association of HVs with LVs that bind to the chromatin is required at a first step during the fusion of vesicles. We found that salt-treated HVs inhibited the binding of LVs to the chromatin. In addition, when salt-treated HVs were pretreated with the proteins in the residue of salt-treatment of HVs, LVs recovered the chromatin-binding activity. In contrast, salt-treatment of LVs did not influence the binding of LVs with chromatin. By using a fluorescence microscopy assay, we showed directly that salt-treated HVs associated with LVs or salt-treated LVs, but HVs did not. These results suggest that soluble mask proteins on HVs keep the two distinct vesicles separate. We propose that the dissociation of mask protein on HVs allows HVs to bind with LVs that are located on the chromatin, subsequently the fusion of each vesicle occurs and the nuclear envelope is formed.

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“…It has been shown that in vitro nuclear assembly spontaneously in Xenopus egg interphase extracts a double nuclear membrane with nuclear pore forms around added chromatin, whether natural animal or plant sperm chromatin or exogenously added procaryotic DNA is used [3][4][5]. Some information concerning reassembly of nuclear membranes, organelles, and individual nucleosomes has arisen from the use of cell-free extracts derived from eggs of Xenopus or sea urchin, and somatic cells and embryos of Drosophila or Nicotiana [2,[6][7][8][9][10][11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

In vitro reassembly of nuclear envelopes and organelles in Xenopus egg extracts

Zheng

Zhai

2006

Cell Res

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We reconstituted bilayer nuclear membranes, multilayer membranes, and organelles from mixtures of Xenopus laevis egg extracts and demembranated Xenopus sperm nuclei. Varying proportions of the cytosolic and vesicular fractions from the eggs were used in the reconstitution mixtures. A cytosol:vesicle ratio of 10:1 promoted reassembly of the normal bilayer nuclear membrane with inserted nuclear pore complexes around the decondensed Xenopus sperm chromatin. A cytosol: vesicle ratio of 5:1 caused decondensed and dispersed sperm chromatin to be either surrounded by or divided by unusual multilayer membrane structures with inlaid pore complexes. A cytosol:vesicle ratio of 2.5:1 promoted reconstitution of mitochondria, endoplasmic reticulum networks, and Golgi apparatus. During reassembly of the endoplasmic reticulum and Golgi apparatus, vesicular fragments of the corresponding organelles fused together and changed their shape to form flattened cisternae, which were then stacked one on top of another.

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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

Cited by 66 publications

References 42 publications

Nuclear Envelope Breakdown Is Coordinated by Both Nup358/RanBP2 and Nup153, Two Nucleoporins with Zinc Finger Modules

Nuclear Envelope Breakdown Is Coordinated by Both Nup358/RanBP2 and Nup153, Two Nucleoporins with Zinc Finger Modules

The interaction of two classes of nuclear vesicles is induced by the dissociation of soluble proteins from one class of vesicles

In vitro reassembly of nuclear envelopes and organelles in Xenopus egg extracts

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