Abstract
Targeting of newly synthesized integral membrane proteins to the appropriate cellular compartment is specified by discrete sequence elements, many of which have been well characterized. An understanding of the signals required to direct integral membrane proteins to the inner nuclear membrane (INM) remains a notable exception. Here we show that integral INM proteins possess basic sequence motifs that resemble ‘classical’ nuclear localization signals. These sequences can mediate direct binding to karyopherin-α and are essential for the passage of integral membrane proteins to the INM. Furthermore, karyopherin-α, karyopherin-β1 and the Ran GTPase cycle are required for INM targeting, underscoring parallels between mechanisms governing the targeting of integral INM proteins and soluble nuclear transport. We also provide evidence that specific nuclear pore complex proteins contribute to this process, suggesting a role for signal-mediated alterations in the nuclear pore complex to allow for passage of INM proteins along the pore membrane.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Gruenbaum, Y., Margalit, A., Goldman, R. D., Shumaker, D. K. & Wilson, K. L. The nuclear lamina comes of age. Nature Rev. Mol. Cell Biol. 6, 21–31 (2005)
Mounkes, L., Kozlov, S., Burke, B. & Stewart, C. L. The laminopathies: nuclear structure meets disease. Curr. Opin. Genet. Dev. 13, 223–230 (2003)
Suntharalingam, M. & Wente, S. R. Peering through the pore: nuclear pore complex structure, assembly, and function. Dev. Cell 4, 775–789 (2003)
Soullam, B. & Worman, H. J. Signals and structural features involved in integral membrane protein targeting to the inner nuclear membrane. J. Cell Biol. 130, 15–27 (1995)
Ohba, T., Schirmer, E. C., Nishimoto, T. & Gerace, L. Energy- and temperature-dependent transport of integral proteins to the inner nuclear membrane via the nuclear pore. J. Cell Biol. 167, 1051–1062 (2004)
Mans, B. J., Anantharaman, V., Aravind, L. & Koonin, E. V. Comparative genomics, evolution and origins of the nuclear envelope and nuclear pore complex. Cell Cycle 3, 1612–1637 (2004)
Rodriguez-Navarro, S., Igual, J. C. & Perez-Ortin, J. E. SRC1: an intron-containing yeast gene involved in sister chromatid segregation. Yeast 19, 43–54 (2002)
Macara, I. G. Transport into and out of the nucleus. Microbiol. Mol. Biol. Rev. 65, 570–594 (2001)
Fried, H. & Kutay, U. Nucleocytoplasmic transport: taking an inventory. Cell. Mol. Life Sci. 60, 1659–1688 (2003)
Weis, K. Regulating access to the genome: nucleocytoplasmic transport throughout the cell cycle. Cell 112, 441–451 (2003)
Kadowaki, T. et al. Isolation and characterization of Saccharomyces cerevisiae mRNA transport-defective (mtr) mutants. J. Cell Biol. 126, 649–659 (1994)
Kalderon, D., Richardson, W. D., Markham, A. F. & Smith, A. E. Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature 311, 33–38 (1984)
Yano, R., Oakes, M. L., Tabb, M. M. & Nomura, M. Yeast Srp1p has homology to armadillo/plakoglobin/β-catenin and participates in apparently multiple nuclear functions including the maintenance of the nucleolar structure. Proc. Natl Acad. Sci. USA 91, 6880–6884 (1994)
Iovine, M. K. & Wente, S. R. A nuclear export signal in Kap95p is required for both recycling the import factor and interaction with the nucleoporin GLFG repeat regions of Nup116p and Nup100p. J. Cell Biol. 137, 797–811 (1997)
Leslie, D. M., Grill, B., Rout, M. P., Wozniak, R. W. & Aitchison, J. D. Kap121p-mediated nuclear import is required for mating and cellular differentiation in yeast. Mol. Cell. Biol. 22, 2544–2555 (2002)
Ryan, K. J. & Wente, S. R. The nuclear pore complex: a protein machine bridging the nucleus and cytoplasm. Curr. Opin. Cell Biol. 12, 361–371 (2000)
Nehrbass, U., Rout, M. P., Maguire, S., Blobel, G. & Wozniak, R. W. The yeast nucleoporin Nup188p interacts genetically and physically with the core structures of the nuclear pore complex. J. Cell Biol. 133, 1153–1162 (1996)
Aitchison, J. D., Rout, M. P., Marelli, M., Blobel, G. & Wozniak, R. W. Two novel related yeast nucleoporins Nup170p and Nup157p: complementation with the vertebrate homologue Nup155p and functional interactions with the yeast nuclear pore-membrane protein Pom152p. J. Cell Biol. 131, 1133–1148 (1995)
Miao, M., Ryan, K. J. & Wente, S. R. The integral membrane protein pom34p functionally links nucleoporin subcomplexes. Genetics 172, 1441–1457 (2006)
Chial, H. J., Rout, M. P., Giddings, T. H. & Winey, M. Saccharomyces cerevisiae Ndc1p is a shared component of nuclear pore complexes and spindle pole bodies. J. Cell Biol. 143, 1789–1800 (1998)
Vaughan, A. et al. Both emerin and lamin C depend on lamin A for localization at the nuclear envelope. J. Cell Sci. 114, 2577–2590 (2001)
Wu, W., Lin, F. & Worman, H. J. Intracellular trafficking of MAN1, an integral protein of the nuclear envelope inner membrane. J. Cell Sci. 115, 1361–1371 (2002)
Ostlund, C., Sullivan, T., Stewart, C. L. & Worman, H. J. Dependence of diffusional mobility of integral inner nuclear membrane proteins on A-type lamins. Biochemistry 45, 1374–1382 (2006)
Horton, P. & Nakai, K. Better prediction of protein cellular localization sites with the k nearest neighbors classifier. Proc. Int. Conf. Intell. Syst. Mol. Biol. 5, 147–152 (1997)
Beilharz, T., Egan, B., Silver, P. A., Hofmann, K. & Lithgow, T. Bipartite signals mediate subcellular targeting of tail-anchored membrane proteins in Saccharomyces cerevisiae. J. Biol. Chem. 278, 8219–8223 (2003)
Booth, J. W., Belanger, K. D., Sannella, M. I. & Davis, L. I. The yeast nucleoporin Nup2p is involved in nuclear export of importin α/Srp1p. J. Biol. Chem. 274, 32360–32367 (1999)
Gilchrist, D., Mykytka, B. & Rexach, M. Accelerating the rate of disassembly of karyopherin·cargo complexes. J. Biol. Chem. 277, 18161–18172 (2002)
Hood, J. K., Casolari, J. M. & Silver, P. A. Nup2p is located on the nuclear side of the nuclear pore complex and coordinates Srp1p/importin-α export. J. Cell Sci. 113, 1471–1480 (2000)
Solsbacher, J., Maurer, P., Vogel, F. & Schlenstedt, G. Nup2p, a yeast nucleoporin, functions in bidirectional transport of importin-α. Mol. Cell. Biol. 20, 8468–8479 (2000)
Saksena, S., Summers, M. D., Burks, J. K., Johnson, A. E. & Braunagel, S. C. Importin-α16 is a translocon-associated protein involved in sorting membrane proteins to the nuclear envelope. Nature Struct. Mol. Biol. 13, 500–508 (2006)
Shulga, N. & Goldfarb, D. S. Binding dynamics of structural nucleoporins govern nuclear pore complex permeability and may mediate channel gating. Mol. Cell. Biol. 23, 534–542 (2003)
Makhnevych, T., Lusk, C. P., Anderson, A. M., Aitchison, J. D. & Wozniak, R. W. Cell cycle regulated transport controlled by alterations in the nuclear pore complex. Cell 115, 813–823 (2003)
Acknowledgements
We are grateful to S. Wente, R. Wozniak, X. Zhao, M. Winey and K. Belanger for yeast strains and plasmids, and M. Rout and J. Novatt for the anti-GFP antibody and Kap95. We also thank A. North for help with the spinning disk confocal microscope, R. Peters and L. Gerace for discussions, and E. Wren for helping us launch this project. We are especially indebted to H. Shio for electron micrograph technical support. This work was supported by an NIH fellowship (to M.C.K.) and the Howard Hughes Medical Institute (to G.B. and C.P.L.).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Competing interests
Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.
Supplementary information
Supplementary Notes
This file contains Supplementary Figures 1–5, Supplementary Methods, and Supplementary Tables 1 and 2. (PDF 3890 kb)
Rights and permissions
About this article
Cite this article
King, M., Lusk, C. & Blobel, G. Karyopherin-mediated import of integral inner nuclear membrane proteins. Nature 442, 1003–1007 (2006). https://doi.org/10.1038/nature05075
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nature05075
This article is cited by
-
ESCRT-III controls nuclear envelope deformation induced by progerin
Scientific Reports (2020)
-
Patrolling the nucleus: inner nuclear membrane-associated degradation
Current Genetics (2019)
-
On the Nuclear Pore Complex and Its Roles in Nucleo-Cytoskeletal Coupling and Mechanobiology
Cellular and Molecular Bioengineering (2016)
-
The tethering of chromatin to the nuclear envelope supports nuclear mechanics
Nature Communications (2015)
-
Subcellular localization of coagulation factor II receptor-like 1 in neurons governs angiogenesis
Nature Medicine (2014)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.