Yeast Nucleoporins Involved in Passive Nuclear Envelope Permeability

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2000 Jun 1

PMID 10831607

Citations 53

Authors

N Shulga

N Mosammaparast

R Wozniak

D S Goldfarb

Affiliations

Soon will be listed here.

Abstract

The vertebrate nuclear pore complex (NPC) harbors an approximately 10-nm diameter diffusion channel that is large enough to admit 50-kD polypeptides. We have analyzed the permeability properties of the Saccharomyces cerevisiae nuclear envelope (NE) using import (NLS) and export (NES) signal-containing green fluorescent protein (GFP) reporters. Compared with wild-type, passive export rates of a classical karyopherin/importin (Kap) Kap60p/Kap95p-targeted NLS-GFP reporter (cNLS-GFP) were significantly faster in nup188-Delta and nup170-Delta cells. Similar results were obtained using two other NLS-GFP reporters, containing either the Kap104p-targeted Nab2p NLS (rgNLS) or the Kap121p-targeted Pho4p NLS (pNLS). Elevated levels of Hsp70 stimulated cNLS-GFP import, but had no effect on the import of rgNLS-GFP. Thus, the role of Hsp70 in NLS-directed import may be NLS- or targeting pathway-specific. Equilibrium sieving limits for the diffusion channel were assessed in vivo using NES-GFP reporters of 36-126 kD and were found to be greater than wild-type in nup188-Delta and nup170-Delta cells. We propose that Nup170p and Nup188p are involved in establishing the functional resting diameter of the NPC's central transport channel.

Citing Articles

Unveiling the complexity: assessing models describing the structure and function of the nuclear pore complex.

Rush C, Jiang Z, Tingey M, Feng F, Yang W Front Cell Dev Biol. 2023; 11:1245939.

PMID: 37876551 PMC: 10591098. DOI: 10.3389/fcell.2023.1245939.

Nucleoporin downregulation modulates progenitor differentiation independent of nuclear pore numbers.

Neely A, Zhang Y, Blumensaadt L, Mao H, Brenner B, Sun C Commun Biol. 2023; 6(1):1033.

PMID: 37853046 PMC: 10584948. DOI: 10.1038/s42003-023-05398-6.

A nuclear pore sub-complex restricts the propagation of Ty retrotransposons by limiting their transcription.

Bonnet A, Chaput C, Palmic N, Palancade B, Lesage P PLoS Genet. 2021; 17(11):e1009889.

PMID: 34723966 PMC: 8585004. DOI: 10.1371/journal.pgen.1009889.

Measuring and Interpreting Nuclear Transport in Neurodegenerative Disease-The Example of C9orf72 ALS.

Semmelink M, Steen A, Veenhoff L Int J Mol Sci. 2021; 22(17).

PMID: 34502125 PMC: 8431710. DOI: 10.3390/ijms22179217.

The Viral Capsid: A Master Key to Access the Host Nucleus.

Blanco-Rodriguez G, Di Nunzio F Viruses. 2021; 13(6).

PMID: 34203080 PMC: 8234750. DOI: 10.3390/v13061178.

References

Danker T, Schillers H, STORCK J, Shahin V, Kramer B, Wilhelmi M . Nuclear hourglass technique: an approach that detects electrically open nuclear pores in Xenopus laevis oocyte. Proc Natl Acad Sci U S A. 1999; 96(23):13530-5. PMC: 23982. DOI: 10.1073/pnas.96.23.13530. View

Adachi M, Fukuda M, Nishida E . Two co-existing mechanisms for nuclear import of MAP kinase: passive diffusion of a monomer and active transport of a dimer. EMBO J. 1999; 18(19):5347-58. PMC: 1171604. DOI: 10.1093/emboj/18.19.5347. View

Kosova B, Pante N, Rollenhagen C, Podtelejnikov A, Mann M, Aebi U . Mlp2p, a component of nuclear pore attached intranuclear filaments, associates with nic96p. J Biol Chem. 2000; 275(1):343-50. DOI: 10.1074/jbc.275.1.343. View

Rout M, AITCHISON J, Suprapto A, Hjertaas K, Zhao Y, Chait B . The yeast nuclear pore complex: composition, architecture, and transport mechanism. J Cell Biol. 2000; 148(4):635-51. PMC: 2169373. DOI: 10.1083/jcb.148.4.635. View

Eddy A, Backen K, Watson G . The concentration of amino acids by yeast cells depleted of adenosine triphosphate. Biochem J. 1970; 120(4):853-8. PMC: 1179680. DOI: 10.1042/bj1200853. View

Paine P, Moore L, Horowitz S . Nuclear envelope permeability. Nature. 1975; 254(5496):109-14. DOI: 10.1038/254109a0. View

Feldherr C, KALLENBACH E, Schultz N . Movement of a karyophilic protein through the nuclear pores of oocytes. J Cell Biol. 1984; 99(6):2216-22. PMC: 2113580. DOI: 10.1083/jcb.99.6.2216. View

Werner-Washburne M, Stone D, Craig E . Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae. Mol Cell Biol. 1987; 7(7):2568-77. PMC: 365392. DOI: 10.1128/mcb.7.7.2568-2577.1987. View

Richardson W, Mills A, Dilworth S, Laskey R, Dingwall C . Nuclear protein migration involves two steps: rapid binding at the nuclear envelope followed by slower translocation through nuclear pores. Cell. 1988; 52(5):655-64. DOI: 10.1016/0092-8674(88)90403-5. View

10.

Nilsson B, Moks T, Jansson B, Abrahmsen L, Elmblad A, Holmgren E . A synthetic IgG-binding domain based on staphylococcal protein A. Protein Eng. 1987; 1(2):107-13. DOI: 10.1093/protein/1.2.107. View

11.

Akey C, Goldfarb D . Protein import through the nuclear pore complex is a multistep process. J Cell Biol. 1989; 109(3):971-82. PMC: 2115752. DOI: 10.1083/jcb.109.3.971. View

12.

Breeuwer M, Goldfarb D . Facilitated nuclear transport of histone H1 and other small nucleophilic proteins. Cell. 1990; 60(6):999-1008. DOI: 10.1016/0092-8674(90)90348-i. View

13.

Reichelt R, Holzenburg A, Buhle Jr E, Jarnik M, Engel A, Aebi U . Correlation between structure and mass distribution of the nuclear pore complex and of distinct pore complex components. J Cell Biol. 1990; 110(4):883-94. PMC: 2116066. DOI: 10.1083/jcb.110.4.883. View

14.

Sherman F . Getting started with yeast. Methods Enzymol. 1991; 194:3-21. DOI: 10.1016/0076-6879(91)94004-v. View

15.

Jarnik M, Aebi U . Toward a more complete 3-D structure of the nuclear pore complex. J Struct Biol. 1991; 107(3):291-308. DOI: 10.1016/1047-8477(91)90054-z. View

16.

Hinshaw J, Carragher B, Milligan R . Architecture and design of the nuclear pore complex. Cell. 1992; 69(7):1133-41. DOI: 10.1016/0092-8674(92)90635-p. View

17.

Okuno Y, Imamoto N, Yoneda Y . 70-kDa heat-shock cognate protein colocalizes with karyophilic proteins into the nucleus during their transport in vitro. Exp Cell Res. 1993; 206(1):134-42. DOI: 10.1006/excr.1993.1129. View

18.

Akey C, Radermacher M . Architecture of the Xenopus nuclear pore complex revealed by three-dimensional cryo-electron microscopy. J Cell Biol. 1993; 122(1):1-19. PMC: 2119598. DOI: 10.1083/jcb.122.1.1. View

19.

Wozniak R, Blobel G, Rout M . POM152 is an integral protein of the pore membrane domain of the yeast nuclear envelope. J Cell Biol. 1994; 125(1):31-42. PMC: 2120016. DOI: 10.1083/jcb.125.1.31. View

20.

Feldherr C, Akin D . Role of nuclear trafficking in regulating cellular activity. Int Rev Cytol. 1994; 151:183-228. DOI: 10.1016/s0074-7696(08)62633-9. View