Regulation of replication timing in fission yeast.
Here we report the first characterization of replication timing and its regulation in the fission yeast Schizosaccharomyces pombe. We used three different synchronization methods: centrifugal elutriation, cdc10 temperature-shift and release, and starvation for deoxyribonucleoside triphosphates (dNTPs) by treatment with hydroxyurea (HU) followed by removal of HU, to study the times when specific autonomously replicating sequence elements (ARS elements; potential replication origins) replicate during S phase. We found that individual ARS elements replicate at characteristic times, some early and some late, independently of synchronization method. In wild-type cells treated with HU, early ARS elements replicated but late ones did not. However, in HU-treated mutant cells lacking the Rad3 (similar to human ATR and ATM) or Cds1 (similar to human CHK2) checkpoint kinase, both early and late ARS elements were able to replicate. Thus under conditions of dNTP starvation the Rad3 and Cds1 kinases are needed to suppress the replication of normally late-replicating regions.
Kim
S M
SM
Department of Cancer Genetics, Roswell Park Cancer Institute, Elm & Carlton Streets, Buffalo, NY 14263, USA.
Huberman
J A
JA
D000251:Adenosine Triphosphatases / Q000172:deficiency / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002498:Centrifugation / Q000379:methods; D064447:Checkpoint Kinase 2; D004265:DNA Helicases / Q000172:deficiency / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects / Q000502:physiology*; D003853:Deoxyribonucleosides / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005434:Flow Cytometry; D018816:Genes, cdc; D006918:Hydroxyurea / Q000494:pharmacology; D009154:Mutation; D011494:Protein Kinases / Q000172:deficiency / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D016196:S Phase / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins; D012568:Schizosaccharomyces / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism; D017384:Sequence Deletion; D013696:Temperature; D013997:Time Factors; D014157:Transcription Factors
D018797:Cell Cycle Proteins; D003853:Deoxyribonucleosides; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; C061178:cdc10 protein, S pombe; D011494:Protein Kinases; D064447:Checkpoint Kinase 2; C578634:CHEK2 protein, human; C578636:Cds1 protein, S pombe; D017346:Protein Serine-Threonine Kinases; D000251:Adenosine Triphosphatases; C061897:Rad3 protein, S cerevisiae; D004265:DNA Helicases; D006918:Hydroxyurea