Nilssen Esben A, Synnes Marianne, Tvegård Tonje, Vebø Heidi, Boye Erik, Grallert Beáta
Department of Cell Biology, Institute for Cancer Research, Montebello, 0310 Oslo, Norway.
BMC Cell Biol. 2004 Oct 21;5(1):40. doi: 10.1186/1471-2121-5-40.
Checkpoint mechanisms prevent cell cycle transitions until previous events have been completed or damaged DNA has been repaired. In fission yeast, checkpoint mechanisms are known to regulate entry into mitosis, but so far no checkpoint inhibiting S phase entry has been identified.
We have studied the response of germinating Schizosaccharomyces pombe spores to UV irradiation in G1. When germinating spores are irradiated in early G1 phase, entry into S phase is delayed. We argue that the observed delay is caused by two separate mechanisms. The first takes place before entry into S phase, does not depend on the checkpoint proteins Rad3, Cds1 and Chk1 and is independent of Cdc2 phosphorylation. Furthermore, it is not dependent upon inhibiting the Cdc10-dependent transcription required for S phase entry, unlike a G1/S checkpoint described in budding yeast. We show that expression of Cdt1, a protein essential for initiation of DNA replication, is delayed upon UV irradiation. The second part of the delay occurs after entry into S phase and depends on Rad3 and Cds1 and is probably due to the intra-S checkpoint. If the germinating spores are irradiated in late G1, they enter S phase without delay and arrest in S phase, suggesting that the delay we observe upon UV irradiation in early G1 is not caused by nonspecific effects of UV irradiation.
We have studied the response of germinating S. pombe spores to UV irradiation in G1 and shown that S phase entry is delayed by a mechanism that is different from classical checkpoint responses. Our results point to a mechanism delaying expression of proteins required for S phase entry.
关卡机制可防止细胞周期转换,直到先前事件完成或受损DNA得到修复。在裂殖酵母中,已知关卡机制可调节进入有丝分裂,但迄今为止尚未发现抑制S期进入的关卡。
我们研究了裂殖酵母孢子在G1期萌发时对紫外线照射的反应。当萌发的孢子在G1早期受到照射时,进入S期会延迟。我们认为观察到的延迟是由两种独立的机制引起的。第一种发生在进入S期之前,不依赖于关卡蛋白Rad3、Cds1和Chk1,且与Cdc2磷酸化无关。此外,与芽殖酵母中描述的G1/S关卡不同,它不依赖于抑制S期进入所需的依赖Cdc10的转录。我们发现,DNA复制起始所必需的蛋白质Cdt1的表达在紫外线照射后会延迟。延迟的第二部分发生在进入S期之后,依赖于Rad3和Cds1,可能是由于S期内关卡。如果萌发的孢子在G1晚期受到照射,它们会毫无延迟地进入S期并在S期停滞,这表明我们在G1早期紫外线照射后观察到的延迟不是由紫外线照射的非特异性效应引起的。
我们研究了裂殖酵母孢子在G1期萌发时对紫外线照射的反应,结果表明进入S期的延迟是由一种不同于经典关卡反应的机制引起的。我们的结果指出了一种延迟S期进入所需蛋白质表达的机制。