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G1-phase and B-type cyclins exclude the DNA-replication factor Mcm4 from the nucleus

Nature Cell Biology volume 1, pages 415–422 (1999)Cite this article

Abstract

Cyclin-dependent kinases (CDKs) activate the firing of replication origins during the S phase of the cell cycle. They also block re-initiation of DNA replication within a single cell cycle, by preventing the assembly of prereplicative complexes at origins. We show here that, in budding yeast, CDKs exclude the essential prereplicative-complex component Mcm4 from the nucleus. Although origin firing can be triggered by the B-type cyclins only, both G1-phase and B-type cyclins cause exit of Mcm4 from the nucleus. These results suggest that G1 cyclins may diminish the cell’s capacity to assemble prereplicative complexes before B-type cyclins trigger origin firing during S phase.

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Figure 1: Mcm4 localization is cell cycle regulated.
Figure 2: Exit of chromatin-bound Mcm4 from the nucleus is dependent upon S-phase progression.
Figure 3: B-cyclin-associated Cdc28 kinase activity blocks nuclear accumulation of Mcm4 in G2/M-arrested cells.
Figure 4: B-cyclins can promote nuclear exit of Mcm4.
Figure 5: Nuclear exit of Mcm4 is activated independently of CLB5,6 expression.
Figure 6: G1-cyclin-dependent nuclear exit of Mcm4.
Figure 7: Cdc6 is required for nuclear accumulation of Mcm4 when Clb–Cdc28 kinases are inactivated in G2/M-arrested cells.
Figure 8: Inactivation of Cdc28 in G2/M-arrested cells is sufficient to cause Cdc6-independent accumulation of Mcm4 in the nucleus.
Figure 9: Regulation of the nuclear localization of Mcm proteins during the budding-yeast cell cycle.

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Acknowledgements

We thank A. Pidoux for suggestions regarding time-lapse analysis of live cells; I. Adams and J. Kilmartin for suggesting the use of a short linker when tagging proteins with GFP; J.A. Tercero for help in growing cultures; K. Boussett, L. Drury, L. Johnston, G. Perkins and A. Varshavsky for plasmids; K. Nasmyth for strains; our colleagues in the Chromosome Replication laboratory and S.E.K.’s group for helpful discussions; and T. Toda for comments on the manuscript. This work was funded by the Imperial Cancer Research Fund, the Cancer Research Campaign, the Wellcome Foundation, and the EU (contract ERB-MRX-CT970125).

Correspondence and requests for materials should be addressed to J.F.X.D.

Supplementary information is available on Nature Cell Biology’s World-Wide Web site (http://cellbio.nature.com ) or as paper copy from the London editorial office of Nature Cell Biology.

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  1. ICRF Clare Hall Laboratories,, South Mimms, Hertfordshire, EN6 3LD, UK

    Karim Labib & John F.X. Diffley

  2. Department of Zoology, South Parks Road, Oxford, OX1 3PS, UK

    Stephen E. Kearsey

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Correspondence to John F.X. Diffley.

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Labib, K., Diffley, J. & Kearsey, S. G1-phase and B-type cyclins exclude the DNA-replication factor Mcm4 from the nucleus. Nat Cell Biol 1, 415–422 (1999). https://doi.org/10.1038/15649

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