elevated levels of the polo kinase cdc5 override the mec1atr checkpoint in budding yeast by acting at different steps of the signaling pathway高浓度的马球激酶cdc5覆盖mec1atr检查点出芽酵母的作用在不同信号通路的步骤.pdfVIP

Elevated Levels of the Polo Kinase Cdc5 Override the Mec1/ATR Checkpoint in Budding Yeast by Acting at Different Steps of the Signaling Pathway 1 1 1 2 Roberto Antonio Donnianni , Matteo Ferrari , Federico Lazzaro , Michela Clerici , Benjamin Tamilselvan 1 1 1 1 Nachimuthu , Paolo Plevani , Marco Muzi-Falconi , Achille Pellicioli * 1 Dipartimento di Scienze Biomolecolari e Biotecnologie, Universita’ degli Studi di Milano, Milano, Italy, 2 Dipartimento di Biotecnologie e Bioscienze, Universita’ di Milano- Bicocca, Milano, Italy Abstract Checkpoints are surveillance mechanisms that constitute a barrier to oncogenesis by preserving genome integrity. Loss of checkpoint function is an early event in tumorigenesis. Polo kinases (Plks) are fundamental regulators of cell cycle progression in all eukaryotes and are frequently overexpressed in tumors. Through their polo box domain, Plks target multiple substrates previously phosphorylated by CDKs and MAPKs. In response to DNA damage, Plks are temporally inhibited in order to maintain the checkpoint-dependent cell cycle block while their activity is required to silence the checkpoint response and resume cell cycle progression. Here, we report that, in budding yeast, overproduction of the Cdc5 polo kinase overrides the checkpoint signaling induced by double strand DNA breaks (DSBs), preventing the phosphorylation of several Mec1/ATR targets, including Ddc2/ATRIP, the checkpoint mediator Rad9, and the transducer kinase Rad53/CHK2. We also show that high levels of Cdc5 slow down DSB processing in a Ra