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The Schizosaccharomyces pombe cdt2(+) gene, a target of G1-S phase-specific transcription factor complex DSC1, is required for mitotic and premeiotic DNA replication
- Source :
- Scopus-Elsevier
- Publication Year :
- 2003
-
Abstract
- We have defined five sev genes by genetic analysis of Schizosaccharomyces pombe mutants, which are defective in both proliferation and sporulation. sev1+/cdt2+ was transcribed during the G1-S phase of the mitotic cell cycle, as well as during the premeiotic S phase. The mitotic expression of cdt2+ was regulated by the MCB-DSC1 system. A mutant of a component of DSC1 affected cdt2+ expression in vivo, and a cdt2+ promoter fragment containing MCB motifs bound DSC1 in vitro. Cdt2 protein also accumulated in S phase and localized to the nucleus. cdt2 null mutants grew slowly at 30° and were unable to grow at 19°. These cdt2 mutants were also medially sensitive to hydroxyurea, camptothecin, and 4-nitroquinoline-1-oxide and were synthetically lethal in combination with DNA replication checkpoint mutations. Flow cytometry analysis and pulsed-field gel electrophoresis revealed that S-phase progression was severely retarded in cdt2 mutants, especially at low temperatures. Under sporulation conditions, premeiotic DNA replication was impaired with meiosis I blocked. Furthermore, overexpression of suc22+, a ribonucleotide reductase gene, fully complemented the sporulation defect of cdt2 mutants and alleviated their growth defect at 19°. These observations suggest that cdt2+ plays an important role in DNA replication in both the mitotic and the meiotic life cycles of fission yeast.
- Subjects :
- DNA Replication
Premeiotic DNA replication
Mutant
Immunoblotting
Molecular Sequence Data
Transcription factor complex
Gene Expression
Mitosis
Electrophoretic Mobility Shift Assay
Biology
Mitotic cell cycle
Schizosaccharomyces
Genetics
Hydroxyurea
Amino Acid Sequence
DNA Primers
DNA replication
Temperature
Sequence Analysis, DNA
biology.organism_classification
Flow Cytometry
Molecular biology
4-Nitroquinoline-1-oxide
Electrophoresis, Gel, Pulsed-Field
Microscopy, Fluorescence
Mutagenesis
Schizosaccharomyces pombe
Camptothecin
Schizosaccharomyces pombe Proteins
Transcription Factors
Research Article
Subjects
Details
- ISSN :
- 00166731
- Volume :
- 164
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Genetics
- Accession number :
- edsair.doi.dedup.....55de4f1b0f9554b8853a4a577a55a9c3