Furnari B, Rhind N, Russell P
Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Science. 1997 Sep 5;277(5331):1495-7. doi: 10.1126/science.277.5331.1495.
Arrest of the cell cycle at the G2 checkpoint, induced by DNA damage, requires inhibitory phosphorylation of the kinase Cdc2 in both fission yeast and human cells. The kinase Wee1 and the phosphatase Cdc25, which regulate Cdc2 phosphorylation, were evaluated as targets of Chk1, a kinase essential for the checkpoint. Fission yeast cdc2-3w Deltacdc25 cells, which express activated Cdc2 and lack Cdc25, were responsive to Wee1 but insensitive to Chk1 and irradiation. Expression of large amounts of Chk1 produced the same phenotype as did loss of the cdc25 gene in cdc2-3w cells. Cdc25 associated with Chk1 in vivo and was phosphorylated when copurified in Chk1 complexes. These findings identify Cdc25, but not Wee1, as a target of the DNA damage checkpoint.
在裂殖酵母和人类细胞中,由DNA损伤诱导的细胞周期在G2期检查点的停滞,需要激酶Cdc2发生抑制性磷酸化。对调节Cdc2磷酸化的激酶Wee1和磷酸酶Cdc25作为检查点所必需的激酶Chk1的作用靶点进行了评估。表达活化Cdc2且缺乏Cdc25的裂殖酵母cdc2-3w Δcdc25细胞对Wee1有反应,但对Chk1和辐射不敏感。在cdc2-3w细胞中,大量表达Chk1产生的表型与缺失cdc25基因相同。Cdc25在体内与Chk1相关联,并且在与Chk1复合物共纯化时会发生磷酸化。这些发现确定Cdc25而非Wee1是DNA损伤检查点的一个靶点。