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J. Biol. Chem., Vol. 276, Issue 48, 44905-44911, November 30, 2001
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From the S-phase onset is controlled, so that it occurs
only once every cell cycle. DNA is licensed for replication after
mitosis in G1, and passage through S-phase
removes the license to replicate. In fission yeast, Cdc6/18 and Cdt1,
two factors required for licensing, are central to ensuring that
replication occurs once per cell cycle. We show that the human
Cdt1 homologue (hCdt1), a nuclear protein, is present only during
G1. After S-phase onset, hCdt1 levels decrease, and it is
hardly detected in cells in early S-phase or G2. hCdt1 can
associate with the DNA replication inhibitor Geminin, however these two
proteins are mostly expressed at different cell cycle stages. hCdt1
mRNA, in contrast to hCdt1 protein, is expressed in
S-phase-arrested cells, and its levels do not change dramatically
during a cell cycle, suggesting that proteolytic rather than
transcriptional controls ensure the timely accumulation of hCdt1.
Consistent with this view, proteasome inhibitors stabilize hCdt1 in
S-phase. In contrast, hCdc6/18 levels are constant through most of the
cell cycle and are only low for a brief period at the end of mitosis.
These results suggest that the presence of active hCdt1 may be crucial
for determining when licensing is legitimate in human cells.
The Human Licensing Factor for DNA Replication Cdt1
Accumulates in G1 and Is Destabilized after Initiation of
S-phase*
§,
Department of Molecular Biology, Graduate
School of Medical Science, Kyushu University, Fukuoka 812-8582, Japan
and the ¶ School of Medicine, University of Patras, 26110 Rio,
Patras, Greece
*
This work was funded by the Ministry of Education, Science
and Culture, by Grants-in-Aid for Specially Promoted Research of Japan,
and by the Association for International Cancer Research (grant 00-271 to Z. L.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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