Differential activation of intra-S-phase checkpoint in response to tripchlorolide and its effects on DNA replication.
DNA replication is tightly regulated during the S phase of the cell cycle, and the activation of the intra-S-phase checkpoint due to DNA damage usually results in arrest of DNA synthesis. However, the molecular details about the correlation between the checkpoint and regulation of DNA replication are still unclear. To investigate the connections between DNA replication and DNA damage checkpoint, a DNA-damage reagent, tripchlorolide, was applied to CHO (Chinese ovary hamster) cells at early- or middle-stages of the S phase. The early-S-phase treatment with TC significantly delayed the progression of the S phase and caused the phosphorylation of the Chk1 checkpoint protein, whereas the middle-S-phase treatment only slightly slowed down the progression of the S phase. Furthermore, the analysis of DNA replication patterns revealed that replication pattern II was greatly prolonged in the cells treated with the drug during the early-S phase, whereas the late-replication patterns of these cells were hardly detected, suggesting that the activation of the intra-S-phase checkpoint inhibits the late-origin firing of DNA replication. We conclude that cells at different stages of the S phase are differentially sensitive to the DNA-damage reagent, and the activation of the intra-S-phase checkpoint blocks the DNA replication progression in the late stage of S phase.
Ren
Yan
Y
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.
Wu
Jia Rui
JR
D000818:Animals; D016466:CHO Cells; D018797:Cell Cycle Proteins / Q000187:drug effects / Q000502:physiology*; D000071877:Checkpoint Kinase 1; D006224:Cricetinae; D004249:DNA Damage / Q000502:physiology; D004261:DNA Replication / Q000187:drug effects* / Q000502:physiology; D004224:Diterpenes / Q000494:pharmacology*; D010616:Phenanthrenes / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism; D016196:S Phase / Q000187:drug effects* / Q000502:physiology*
D018797:Cell Cycle Proteins; D004224:Diterpenes; D010616:Phenanthrenes; C068051:tripchlorolide; D011494:Protein Kinases; D000071877:Checkpoint Kinase 1