Nucleocytoplasmic Transport

Görlich, Dirk; Jäkel, Stefan

doi:10.1016/b978-012200731-6.50015-x

Cited by 114 publications

(157 citation statements)

References 172 publications

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“…The partial nuclear localization of GFP is caused by bidirectional diffusion through the nuclear pore complex (von Arnim et al, 1998). The exclusion limit for this bidirectional diffusion has been estimated to 40 to 60 kD (Gorlich and Mattaj, 1996). The predicted size of the GFP-AtPER1 fusion protein of 52 kD underpasses the size exclusion limit, so we cannot exclude that the partial nuclear localization seen for GFP-AtPER1 is due to diffusion.…”

Section: Discussionmentioning

confidence: 99%

Seed 1-Cysteine Peroxiredoxin Antioxidants Are Not Involved in Dormancy, But Contribute to Inhibition of Germination during Stress

et al. 2003

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Peroxiredoxins (Prx) are thiol-dependent antioxidants containing one (1-cysteine [-Cys]) or two (2-Cys) conserved Cys residues that protect lipids, enzymes, and DNA against reactive oxygen species. In plants, the 1-Cys Prxs are highly expressed during late seed development, and the expression pattern is dormancy related in mature seeds. We have expressed the Arabidopsis 1-Cys Prx AtPER1 in Escherichia coli and show that this protein has antioxidant activity in vitro and protects E. coli in vivo against the toxic oxidant cumene hydroperoxide. Although some 1-Cys Prxs are targeted to the nucleus, a green fluorescent protein-AtPER1 fusion protein was also localized to the cytoplasm in an onion epidermis subcellular localization assay. It has been proposed that seed Prxs are involved in maintenance of dormancy and/or protect the embryo and aleurone layer surviving desiccation against damage caused by reactive oxygen species. These hypotheses were tested using transgenic Arabidopsis lines overexpressing the barley (Hordeum vulgare) 1-Cys PER1 protein and lines with reduced levels of AtPER1 due to antisensing or RNA interference. We found no correlation between Prx levels and the duration of the after-ripening period required before germination. Thus, Prxs are unlikely to contribute to maintenance of dormancy. RNA interference lines almost devoid of AtPER1 protein developed and germinated normally under standard growth room conditions. However, seeds from lines overexpressing PER1 were less inclined to germinate than wild-type seeds in the presence of NaCl, mannitol, and methyl viologen, suggesting that Prx can sense harsh environmental surroundings and play a part in the inhibition of germination under unfavorable conditions.

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Section: Discussionmentioning

confidence: 99%

Seed 1-Cysteine Peroxiredoxin Antioxidants Are Not Involved in Dormancy, But Contribute to Inhibition of Germination during Stress

et al. 2003

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“…The NES is a short leucine-rich sequence motif that binds Crm-1 and is necessary to mediate active nuclear export (Gorlich and Mattaj, 1996). NES transport motifs regulate the intracellular localization of a variety of proteins (Xu and Massague, 2004).…”

Section: Role Of the Regulatory Domains For Nucleocytoplasmic Shuttlimentioning

confidence: 99%

Role of the Regulatory Domain of Protein Kinase D2 in Phorbol Ester Binding, Catalytic Activity, and Nucleocytoplasmic Shuttling

Auer

Blume

Sturany

et al. 2005

MBoC

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Protein kinase D2 (PKD2) belongs to the PKD family of serine/threonine kinases that is activated by phorbol esters and G protein-coupled receptors (GPCRs). Its C-terminal regulatory domain comprises two cysteine-rich domains (C1a/C1b) followed by a pleckstrin homology (PH) domain. Here, we examined the role of the regulatory domain in PKD2 phorbol ester binding, catalytic activity, and subcellular localization: The PH domain is a negative regulator of kinase activity. C1a/C1b, in particular C1b, is required for phorbol ester binding and gastrin-stimulated PKD2 activation, but it has no inhibitory effect on the catalytic activity. Gastrin triggers nuclear accumulation of PKD2 in living AGS-B cancer cells. C1a/C1b, not the PH domain, plays a complex role in the regulation of nucleocytoplasmic shuttling: We identified a nuclear localization sequence in the linker region between C1a and C1b and a nuclear export signal in the C1a domain. In conclusion, our results define the critical components of the PKD2 regulatory domain controlling phorbol ester binding, catalytic activity, and nucleocytoplasmic shuttling and reveal marked differences to the regulatory properties of this domain in PKD1. These findings could explain functional differences between PKD isoforms and point to a functional role of PKD2 in the nucleus upon activation by GPCRs.

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“…Finally, we tested the substrate proteins for their binding to importin-b, which is a key transporter that promotes transportation of cargo proteins across the p53 nuclear importation Q Li et al nuclear membrane (Gorlich and Mattaj, 1996). GST-pulldown showed that importin-b coprecipitated with the GST-p53BNLS-EGFP protein, but not GST-p53MNLS-EGFP (Figure 1d), suggesting that the lack of nuclear import activity of the monopartite p53 NLS is due to its inability to bind importin-b.…”

Section: Resultsmentioning

confidence: 99%

“…However, two additional amino acids located 5 0 of the original NLSI were showed to be required to form a fully functional NLS, indicating that the p53 NLS was bipartite in nature (Liang and Clarke, 1999). Nuclear import is initialized by binding of importin-a/b complex, which mediates the docking of proteins at the nuclear pore complex (Gorlich and Mattaj, 1996). However, the mechanism that regulates the importin-mediated nuclear import is still poorly understood.…”

Section: Introductionmentioning

confidence: 99%

Genetic analysis of p53 nuclear importation

Falsey

Gaitonde

et al. 2007

Oncogene

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A key step in activation of the p53 tumor suppressor is its transport into the nucleus; however, despite intensive study of p53, the regulation of its subcellular localization is still poorly understood. Here we examined the p53 nuclear importation using a series of mutant cell lines that were resistant to the growth inhibitory effects of temperaturesensitive murine p53 (tsp53). Examination of the p53 subcellular localization in these cell lines showed that the protein was cytoplasmic in most of them. Using a digitoninpermeabilized cell in vitro nuclear import system, we show that cytosols from these cell lines do not support nuclear translocation of a p53 nuclear localization signal (NLS)-containing substrate protein, but promote nuclear localization of a SV40TAgNLS-containing substrate. Complementation assays and use of the mutant cells themselves in the in vitro assays demonstrate that both soluble and insoluble protein components are involved in p53 nuclear import. Collectively, our results suggest that there is a p53 NLS-selective nuclear import pathway and that both soluble and insoluble proteins are involved in its function.

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Nucleocytoplasmic Transport

Cited by 114 publications

References 172 publications

Seed 1-Cysteine Peroxiredoxin Antioxidants Are Not Involved in Dormancy, But Contribute to Inhibition of Germination during Stress

Seed 1-Cysteine Peroxiredoxin Antioxidants Are Not Involved in Dormancy, But Contribute to Inhibition of Germination during Stress

Role of the Regulatory Domain of Protein Kinase D2 in Phorbol Ester Binding, Catalytic Activity, and Nucleocytoplasmic Shuttling

Genetic analysis of p53 nuclear importation

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