Nuclear volume and the number of nuclear pore complexes (NPCs) on the nucleus almost double during interphase in dividing cells. How these events are coordinated with the cell cycle is poorly understood, particularly in mammalian cells. We report here, based on newly developed techniques for visualizing NPC formation, that cyclin-dependent kinases (Cdks), especially Cdk1 and Cdk2, promote interphase NPC formation in human dividing cells. Cdks seem to drive an early step of NPC formation because Cdk inhibition suppressed generation of 'nascent pores', which we argue are immature NPCs under the formation process. Consistent with this, Cdk inhibition disturbed proper expression and localization of some nucleoporins, including Elys/Mel-28, which triggers postmitotic NPC assembly. Strikingly, Cdk suppression did not notably affect nuclear growth, suggesting that interphase NPC formation and nuclear growth have distinct regulation mechanisms.
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Morgan, D.O. The Cell Cycle—Principles of Control. (New Science Press Ltd, London, 2007).
Hetzer, M.W., Walther, T.C. & Mattaj, I.W. Pushing the envelope: structure, function, and dynamics of the nuclear periphery. Annu. Rev. Cell Dev. Biol. 21, 347–380 (2005).
Tran, E.J. & Wente, S.R. Dynamic nuclear pore complexes: life on the edge. Cell 125, 1041–1053 (2006).
Antonin, W., Ellenberg, J. & Dultz, E. Nuclear pore complex assembly through the cell cycle: regulation and membrane organization. FEBS Lett. 582, 2004–2016 (2008).
D'Angelo, M.A. & Hetzer, M.W. Structure, dynamics and function of nuclear pore complexes. Trends Cell Biol. 18, 456–466 (2008).
Lim, R.Y., Ullman, K.S. & Fahrenkrog, B. Biology and biophysics of the nuclear pore complex and its components. Int. Rev. Cell. Mol. Biol. 267, 299–342 (2008).
Harel, A. et al. Removal of a single pore subcomplex results in vertebrate nuclei devoid of nuclear pores. Mol. Cell 11, 853–864 (2003).
Walther, T.C. et al. The conserved Nup107–160 complex is critical for nuclear pore complex assembly. Cell 113, 195–206 (2003).
Rabut, G., Lenart, P. & Ellenberg, J. Dynamics of nuclear pore complex organization through the cell cycle. Curr. Opin. Cell Biol. 16, 314–321 (2004).
D'Angelo, M.A., Raices, M., Panowski, S.H. & Hetzer, M.W. Age-dependent deterioration of nuclear pore complexes causes a loss of nuclear integrity in postmitotic cells. Cell 136, 284–295 (2009).
Maul, G.G. et al. Time sequence of nuclear pore formation in phytohemagglutinin-stimulated lymphocytes and in HeLa cells during the cell cycle. J. Cell Biol. 55, 433–447 (1972).
Maul, G.G. Nuclear pore complexes. Elimination and reconstruction during mitosis. J. Cell Biol. 74, 492–500 (1977).
Onischenko, E.A., Gubanova, N.V., Kiseleva, E.V. & Hallberg, E. Cdk1 and okadaic acid-sensitive phosphatases control assembly of nuclear pore complexes in Drosophila embryos. Mol. Biol. Cell 16, 5152–5162 (2005).
Bodoor, K. et al. Sequential recruitment of NPC proteins to the nuclear periphery at the end of mitosis. J. Cell Sci. 112, 2253–2264 (1999).
Dultz, E. et al. Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells. J. Cell Biol. 180, 857–865 (2008).
Rasala, B.A., Orjalo, A.V., Shen, Z., Briggs, S. & Forbes, D.J. ELYS is a dual nucleoporin/kinetochore protein required for nuclear pore assembly and proper cell division. Proc. Natl. Acad. Sci. USA 103, 17801–17806 (2006).
Franz, C. et al. MEL-28/ELYS is required for the recruitment of nucleoporins to chromatin and postmitotic nuclear pore complex assembly. EMBO Rep. 8, 165–172 (2007).
Rasala, B.A., Ramos, C., Harel, A. & Forbes, D.J. Capture of AT-rich chromatin by ELYS recruits POM121 and NDC1 to initiate nuclear pore assembly. Mol. Biol. Cell 19, 3982–3996 (2008).
Walther, T.C. et al. RanGTP mediates nuclear pore complex assembly. Nature 424, 689–694 (2003).
D'Angelo, M.A., Anderson, D.J., Richard, E. & Hetzer, M.W. Nuclear pores form de novo from both sides of the nuclear envelope. Science 312, 440–443 (2006).
Maeshima, K. et al. Cell-cycle-dependent dynamics of nuclear pores: pore-free islands and lamins. J. Cell Sci. 119, 4442–4451 (2006).
Bach, S. et al. Roscovitine targets, protein kinases and pyridoxal kinase. J. Biol. Chem. 280, 31208–31219 (2005).
Whittaker, S.R. et al. The cyclin-dependent kinase inhibitor seliciclib (R-roscovitine; CYC202) decreases the expression of mitotic control genes and prevents entry into mitosis. Cell Cycle 6, 3114–3131 (2007).
Ikegami, S. et al. Aphidicolin prevents mitotic cell division by interfering with the activity of DNA polymerase-α. Nature 275, 458–460 (1978).
Pedrali-Noy, G. et al. Synchronization of HeLa cell cultures by inhibition of DNA polymerase α with aphidicolin. Nucleic Acids Res. 8, 377–387 (1980).
Maul, H.M., Hsu, B.Y., Borun, T.M. & Maul, G.G. Effect of metabolic inhibitors on nuclear pore formation during the HeLa S3 cell cycle. J. Cell Biol. 59, 669–676 (1973).
Wang, J.C. Cellular roles of DNA topoisomerases: a molecular perspective. Nat. Rev. Mol. Cell Biol. 3, 430–440 (2002).
Kill, I.R. Localisation of the Ki-67 antigen within the nucleolus. Evidence for a fibrillarin-deficient region of the dense fibrillar component. J. Cell Sci. 109, 1253–1263 (1996).
Leung, A.K. & Lamond, A.I. The dynamics of the nucleolus. Crit. Rev. Eukaryot. Gene Expr. 13, 39–54 (2003).
Nagai, T. et al. A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nat. Biotechnol. 20, 87–90 (2002).
Rekas, A., Alattia, J.R., Nagai, T., Miyawaki, A. & Ikura, M. Crystal structure of venus, a yellow fluorescent protein with improved maturation and reduced environmental sensitivity. J. Biol. Chem. 277, 50573–50578 (2002).
Katahira, J. et al. The Mex67p-mediated nuclear mRNA export pathway is conserved from yeast to human. EMBO J. 18, 2593–2609 (1999).
Rabut, G., Doye, V. & Ellenberg, J. Mapping the dynamic organization of the nuclear pore complex inside single living cells. Nat. Cell Biol. 6, 1114–1121 (2004).
Gray, N.S. et al. Exploiting chemical libraries, structure, and genomics in the search for kinase inhibitors. Science 281, 533–538 (1998).
Iino, H. et al. Live imaging system for visualizing nuclear pore complex (NPC) formation during interphase in mammalian cells. Genes Cells 15, 647–660 (2010).
L'Italien, L., Tanudji, M., Russell, L. & Schebye, X.M. Unmasking the redundancy between Cdk1 and Cdk2 at G2 phase in human cancer cell lines. Cell Cycle 5, 984–993 (2006).
Skoufias, D.A., Indorato, R.L., Lacroix, F., Panopoulos, A. & Margolis, R.L. Mitosis persists in the absence of Cdk1 activity when proteolysis or protein phosphatase activity is suppressed. J. Cell Biol. 179, 671–685 (2007).
Dudley, D.T., Pang, L., Decker, S.J., Bridges, A.J. & Saltiel, A.R. A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc. Natl. Acad. Sci. USA 92, 7686–7689 (1995).
Hawryluk-Gara, L.A., Shibuya, E.K. & Wozniak, R.W. Vertebrate Nup53 interacts with the nuclear lamina and is required for the assembly of a Nup93-containing complex. Mol. Biol. Cell 16, 2382–2394 (2005).
Hawryluk-Gara, L.A., Platani, M., Santarella, R., Wozniak, R.W. & Mattaj, I.W. Nup53 is required for nuclear envelope and nuclear pore complex assembly. Mol. Biol. Cell 19, 1753–1762 (2008).
Neumann, F.R. & Nurse, P. Nuclear size control in fission yeast. J. Cell Biol. 179, 593–600 (2007).
Jorgensen, P. et al. The size of the nucleus increases as yeast cells grow. Mol. Biol. Cell 18, 3523–3532 (2007).
Huber, M.D. & Gerace, L. The size-wise nucleus: nuclear volume control in eukaryotes. J. Cell Biol. 179, 583–584 (2007).
Zink, D., Fischer, A.H. & Nickerson, J.A. Nuclear structure in cancer cells. Nat. Rev. Cancer 4, 677–687 (2004).
Ohashi, M. et al. A new human diploid cell strain, TIG-1, for the research on cellular aging. Exp. Gerontol. 15, 121–133 (1980).
Yahata, K. et al. cHS4 insulator-mediated alleviation of promoter interference during cell-based expression of tandemly associated transgenes. J. Mol. Biol. 374, 580–590 (2007).
Maeshima, K. & Laemmli, U.K. A two-step scaffolding model for mitotic chromosome assembly. Dev. Cell 4, 467–480 (2003).
Peranen, H., Rikkonen, M. & Kaariainen, L. A method for exposing hidden antigenic sites in paraformaldehyde-fixed cultured cells, applied to initially unreactive antibodies. J. Histochem. Cytochem. 41, 447–454 (1993).
We are grateful to K. Hamasuna and Y. Sasaki for competent technical assistance in this study; K. Wilson (Johns Hopkins Univ.), A. Miyawaki (RIKEN), T. Nagai (Hokkaido Univ.), R. Tsien (Univ. of California, San Diego), T. Tachibana (Osaka City Univ.), V. Doye (Institut Jacques Monod), G. Felsenfeld (US National Institutes of Health), B. Burke (Institute of Medical Biology) and M. Takagi (RIKEN) for their generous gifts of materials; M. Hiroshima, T. Haraguchi, H. Araki and members of the Cellular Dynamics Lab at RIKEN for helpful discussions; and H. Niki for the access to DeltaVision microscope at the Japanese National Institute of Genetics. This work was supported by a Japanese Ministry of Education, Culture, Sports, Science and Technology grant-in-aid, by RIKEN Special Project Funding for Basic Science (Bioarchitect Project) and by RIKEN R&D (President's Discretionary Fund).
The authors declare no competing financial interests.
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Maeshima, K., Iino, H., Hihara, S. et al. Nuclear pore formation but not nuclear growth is governed by cyclin-dependent kinases (Cdks) during interphase. Nat Struct Mol Biol 17, 1065–1071 (2010). https://doi.org/10.1038/nsmb.1878
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