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Science 22 December 2000:
Vol. 290. no. 5500, pp. 2309 - 2312
DOI: 10.1126/science.290.5500.2309
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Reports

Inhibition of Eukaryotic DNA Replication by Geminin Binding to Cdt1

James A. Wohlschlegel,1 Brian T. Dwyer,1 Suman K. Dhar,1 Christin Cvetic,2 Johannes C. Walter,2 Anindya Dutta1*

In all eukaryotic organisms, inappropriate firing of replication origins during the G2 phase of the cell cycle is suppressed by cyclin-dependent kinases. Multicellular eukaryotes contain a second putative inhibitor of re-replication called geminin. Geminin is believed to block binding of the mini-chromosome maintenance (MCM) complex to origins of replication, but the mechanism of this inhibition is unclear. Here we show that geminin interacts tightly with Cdt1, a recently identified replication initiation factor necessary for MCM loading. The inhibition of DNA replication by geminin that is observed in cell-free DNA replication extracts is reversed by the addition of excess Cdt1. In the normal cell cycle, Cdt1 is present only in G1 and S, whereas geminin is present in S and G2 phases of the cell cycle. Together, these results suggest that geminin inhibits inappropriate origin firing by targeting Cdt1.

1 Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, 75 Francis Street, Boston, MA 02115, USA.
2 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
*   To whom correspondence should be addressed. E-mail: aduttta{at}rics.bwh.harvard.edu

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