Nuclear protein migration involves two steps: Rapid binding at the nuclear envelope followed by slower translocation through nuclear pores

Richardson, William D.; Mills, Anthony D.; Dilworth, S. J.; Laskey, Ronald A.; Dingwall, Colin

doi:10.1016/0092-8674(88)90403-5

Cited by 525 publications

(369 citation statements)

References 29 publications

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“…Since active translocation across nuclear pores requires energy 25,26 , nuclear proteins that are not retained should diffuse out of the nucleus when ATP is depleted 27 . We depleted ATP from HeLa cells by glucose deprivation and assessed the localization of AID L198S-GFP, a constitutively nuclear AID variant carrying a point mutation inactivating the NES 18 , and NLS SV40 -GFP.…”

Section: Aid Is Actively Imported Into the Nucleusmentioning

confidence: 99%

Active nuclear import and cytoplasmic retention of activation-induced deaminase

Patenaude

Orthwein

et al. 2009

Nat Struct Mol Biol

133

175

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The enzyme Activation Induced Deaminase (AID) triggers antibody diversification in B-cells by catalyzing deamination and consequently mutation of immunoglobulin genes. To minimize off-target deamination, AID is restrained by several regulatory mechanisms including nuclear exclusion, thought to be mediated exclusively by active nuclear export. Here we identify two other mechanisms involved in controlling AID subcellular localization. AID is unable to passively diffuse into the nucleus, despite its small size, its nuclear entry requiring active import mediated by a conformational nuclear localization sequence (NLS). We also identify a determinant for AID cytoplasmic retention in its Cterminus, which hampers diffusion to the nucleus, competes with nuclear import and is critical for maintaining the predominantly cytoplasmic localization of AID in steady-state conditions. Blocking nuclear import alters the balance between these processes in favor of cytoplasmic retention, resulting in reduced isotype class switching.3

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Section: Aid Is Actively Imported Into the Nucleusmentioning

confidence: 99%

Active nuclear import and cytoplasmic retention of activation-induced deaminase

Patenaude

Orthwein

et al. 2009

Nat Struct Mol Biol

133

175

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“…The import of proteins into the nucleus consists of at least two steps. The first is a NLS-dependent binding of the nuclear import substrate to the cytoplasmic side of the nuclear pore complex followed by an ATP-dependent translocation through the pore [5,6,8,9].…”

Section: Introductionmentioning

confidence: 99%

Nuclear protein transport is functionally conserved between yeast and higher eukaryotes

Wagner

Hall

1993

FEBS Letters

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The ability of a yeast nuclear protein to be transported into the nucleus of a higher eukaryotic cell was investigated. Mcml, a transcriptional activator protein from Succharomyces cerevisiue, was microinjected into the cytoplasm of Xenopus Iuevis frog oocytes. Mcml was imported into the oocyte nucleus indicating that the machinery for nuclear transport is conserved from yeast to higher eukaryotes. Furthermore, by comparing the nuclear import of free proteins and protein-gold complexes, we found that protein-gold complex formation appears to partially and specifically inactivate the nuclear transport activity of Mcml and that nucleoplasmin is an exceptionally good nuclear import substrate.

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“…Three-dimensional image reconstruction suggests that they are comprised of nuclear and cytoplasmic rings, with eight spokes radiating inward toward a central transporter apparatus (Unwin and Milligan, 1982;Akey, 1990Akey, , 1991Reichelt et al, 1990;Jarnik and Aebi, 1991;Hinshaw et al, 1992;Akey and Radermacher, 1993). Under certain preparation conditions, electron microscopy permits filaments to be observed projecting from NPCs into both the nucleoplasm and cytoplasm (Maul, 1977;Richardson et al, 1988;Allen and Douglas, 1989;Jarnik and Aebi, 1991;Ris and Malecki, 1993). The nucleoplasmic filaments sometimes appear organized into basket-like structures, termed the fishtrap (arnik and Aebi, 1991;Goldberg and Allen, 1992).…”

mentioning

confidence: 99%

Mutation or deletion of the Saccharomyces cerevisiae RAT3/NUP133 gene causes temperature-dependent nuclear accumulation of poly(A)+ RNA and constitutive clustering of nuclear pore complexes.

Heath

Amberg

et al. 1995

MBoC

119

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To identify genes whose products play potential roles in the nucleocytoplasmic export of messenger RNA, we isolated temperature-sensitive strains of Saccharomyces cerevisiae and examined them by fluorescent in situ hybridization. With the use of a digoxigen-tagged oligo-(dT)50 probe, we identified those that showed nuclear accumulation of poly(A)+ RNA when cells were shifted to the nonpermissive temperature. We describe here the properties of yeast strains bearing the rat3-1 mutation (RAT -ribonucleic acid trafficking) and the cloning of the RAT3 gene. When cultured at the permissive temperature of 23°C, fewer than 10% of cells carrying the rat3-1 allele showed nuclear accumulation of poly(A)+ RNA, whereas approximately 70% showed nuclear accumulation of poly(A)+ RNA after a shift to 37°C for 4 h. In wild-type cells, nuclear pore complexes (NPCs) are distributed relatively evenly around the nuclear envelope. Both indirect immunofluorescence analysis and electron microscopy of rat3-1 cells indicated that NPCs were clustered into one or a few regions of the NE in mutant cells. Similar NPC clustering was seen in mutant cells cultured at temperatures between 15°C and 37°C. The RAT3 gene encodes an 1157-amino acid protein without similarity to other known proteins. It is essential for growth only at 37°C. Cells carrying a disruption of the RAT3 gene were very similar to cells carrying the original rat3-1 mutation; they showed temperature-dependent nuclear accumulation of poly(A)+ RNA and exhibited constitutive clustering of NPCs. Epitope tagging of Rat3p demonstrated that it is located at the nuclear periphery and co-localizes with nuclear pore proteins recognized by the RL1 monoclonal antibody. We refer to this nucleoporin as Rat3p/Nupl33p.

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Nuclear protein migration involves two steps: Rapid binding at the nuclear envelope followed by slower translocation through nuclear pores

Cited by 525 publications

References 29 publications

Active nuclear import and cytoplasmic retention of activation-induced deaminase

Active nuclear import and cytoplasmic retention of activation-induced deaminase

Nuclear protein transport is functionally conserved between yeast and higher eukaryotes

Mutation or deletion of the Saccharomyces cerevisiae RAT3/NUP133 gene causes temperature-dependent nuclear accumulation of poly(A)+ RNA and constitutive clustering of nuclear pore complexes.

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