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Germinating fission yeast spores delay in G1 in response to UV irradiation
Esben A Nilssen1, Marianne Synnes, Tonje Tvegård
1Department of Cell Biology, Institute for Cancer Research, Montebello, 0310 Oslo, Norway. m18@online.no <m18@online.no>
Background:
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.
Results:
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.
Conclusions:
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.
Insights
UV irradiation in G1 delays S phase entry in fission yeast via a novel mechanism. This process involves delayed expression of proteins essential for DNA replication initiation, distinct from known checkpoint pathways.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Checkpoint mechanisms ensure cell cycle completion before transitions.
- Fission yeast (Schizosaccharomyces pombe) has known mitotic checkpoints, but none inhibiting S phase entry.
- Understanding G1/S regulation is crucial for cell cycle control.
Purpose of the Study:
- To investigate the response of S. pombe spores to UV irradiation during G1 phase.
- To identify novel checkpoints regulating entry into S phase.
- To elucidate the mechanisms underlying UV-induced delays in S phase entry.
Main Methods:
- Irradiation of germinating S. pombe spores in early and late G1.
- Analysis of S phase entry and cell cycle progression.
- Investigation of checkpoint protein involvement (Rad3, Cds1, Chk1).
- Assessment of Cdc2 phosphorylation and Cdc10-dependent transcription.
- Monitoring of Cdt1 expression, a key protein for DNA replication.
Main Results:
- UV irradiation in early G1 delays S phase entry.
- This delay involves two mechanisms: one pre-S phase independent of known checkpoint proteins and Cdc2 phosphorylation, and another intra-S phase dependent on Rad3 and Cds1.
- UV irradiation delays the expression of Cdt1, essential for DNA replication initiation.
- Irradiation in late G1 leads to immediate S phase entry and subsequent arrest, ruling out nonspecific UV effects.
Conclusions:
- S. pombe spores exhibit a novel mechanism delaying S phase entry upon UV irradiation in G1.
- This mechanism differs from classical checkpoint responses and involves delayed expression of S phase entry proteins.
- The findings reveal a new layer of cell cycle regulation in response to DNA damage.
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