Nuclear pore formation but not nuclear growth is governed by cyclin-dependent kinases (Cdks) during interphase

Abstract

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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Figure 1: NPC density and nuclear volume increase during the cell cycle.
Figure 2: Heterokaryon technique for visualization of NPC formation.
Figure 3: Cdk-dependent NPC formation during interphase.
Figure 4: Cdk-dependent NPC distribution and direct observation of NPC structures.
Figure 5: Cdks control expression and localization of several nucleoporins.
Figure 6: Cdk activity is not required for postmitotic NPC assembly.

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Acknowledgements

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).

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K.M. designed the experiments; K.M., H.I. and S.H. performed most of the experiments; A.W. assisted with some experiments; R.N., K.M. and T.H. performed cryo-SEM observations; M.N., K.M. and H.Y. carried out quantitative analyses; T.F., K.Y. and F.I. made some materials; N.I. advised throughout the study; K.M. and N.I. wrote the paper.

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Correspondence to Kazuhiro Maeshima or Naoko Imamoto.

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The authors declare no competing financial interests.

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Supplementary Figures 1–7 and Supplementary Methods (PDF 9392 kb)

Supplementary Video 1

Time-lapse movie of HeLa cells stably expressing H2B–mRFP1 and EGFP (MOV 4058 kb)

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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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