Ribonucleotide reductase activity during the cell cycle of Saccharomyces cerevisiae

Lowdon, Margaret; Vitols, E.

doi:10.1016/0003-9861(73)90611-5

Cited by 68 publications

(31 citation statements)

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“…The enzymatic activity of ribonucleotide reductase fluctuates in the cell cycle with an activity maximum in S phase in all eukaryotic cells that have been examined, including yeast (Lowden and Vitols 1973). In mammalian cells, this regulation is accomplished by modulating the levels of the small subunit, whereas the large subunit remains constant throughout the cell cycle (Engstrom et al 19851.…”

Section: Transcription Of Rnr1 Is Tightly Cell-cycle Regulatedmentioning

confidence: 99%

“…romyces cerevisiae, not only are genes encoding the enzymatic machinery for DNA synthesis cell-cycle regulated [for example POLl, DNA polymerase I (Johnston et al 1987); CDC9, DNA ligase (Barker et al 1985;Peterson et al 1985)], but so are many of the enzymatic activities involved in the production of the dNTP precursors needed for DNA synthesis [for example, CDC8, thymidylate kinase (White et al 1988); CDC21, thymidylate synthase IStorms et al 1984) and ribonucleotide reductase (Lowden and Vitols 1973)]. Several other genes associated with DNA metabolism are also cell-cycle regulated; these include histones (Hereford et al 1981); HO, an endonuclease involved in mating type switching; SW15, a regulator of mating type switching (Nasmyth 1987); and RAD6 (Kupiec and Simchen 1986).…”

mentioning

confidence: 99%

“…In yeast, the small subunit is encoded by RNR2 (Elledge and Davis 1987;Hurd et al 1987). The activity of the enzyme fluctuates in the cell cycle with a maximum in S phase (Lowden and Vitols 1973). The amount of ribonucleotide reductase is also increased under conditions of nucleotide depletion (Lammers and Follman 1984).…”

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confidence: 99%

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Two genes differentially regulated in the cell cycle and by DNA-damaging agents encode alternative regulatory subunits of ribonucleotide reductase.

Elledge¹,

Davis²

1990

Genes Dev.

284

234

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Ribonucleotide reductase activity is essential for progression through the cell cycle, catalyzing the rate-limiting step for the production of deoxyribonucleotides needed for DNA synthesis. The enzymatic activity of the enzyme fluctuates in the cell cycle with an activity maximum in S phase. We have identified and characterized two Saccharomyces cerevisiae genes encoding the regulatory subnnit of ribonucleotide reductase, RNR1 and RNR3. They share -80% amino acid identity with each other and 60% with the mammalian homolog, MI. Genetic disruption reveals that the RNR1 gene is essential for mitotic viability, whereas the RNR3 gene is not essential. A high-copy-number clone of RNR3 is able to suppress the lethality of rarl mutations. Analysis of mRNA levels in cell-cycle-synchronized cultures reveals that the RNR1 mRNA is tightly cell-cycle regulated, fluctuating 15-to 30-fold, and is coordinately regulated with the POLl mRNA, being expressed in the late Gz and S phases of the cell cycle. Progression from the a-factor-induced G~ block to induction of RNR1 mRNA is blocked by cycloheximide, further defining the requirement for protein synthesis in the G~-to S-phase transition. Both RNR1 and RNR3 transcripts are inducible by treatments that damage DNA, such as 4-nitroquinoline-l-oxide and methylmethanesulfonate, or block DNA replication, such as hydroxyurea. RNR1 is inducible 3-to 5-fold, and RNR3 is inducible > 100-fold. When MATa ceils are arrested in GI by a-factor, RNR1 and RNR3 mRNA is still inducible by DNA damage, indicating that the observed induction can occur outside of S phase. Inhibition of ribonucleotide reductase activity by hydroxyurea treatment results in arrest of the cell cycle in S phase as large budded, uninucleate cells. This specific cell-cycle arrest is independent of the RAD9 gene, defining a separate pathway for the coordination of DNA synthesis and cell-cycle progression.

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Section: Transcription Of Rnr1 Is Tightly Cell-cycle Regulatedmentioning

confidence: 99%

mentioning

confidence: 99%

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Two genes differentially regulated in the cell cycle and by DNA-damaging agents encode alternative regulatory subunits of ribonucleotide reductase.

Elledge¹,

Davis²

1990

Genes Dev.

284

234

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“…It is the first consistent demonstration of the NDP --* + dNTP + DNA pathway in plant cells. In fact, these parameters coincide more precisely during the Scenedesmus cell cycle than in any other organism studied, including yeast [25] or phytohemagglutininstimulated lymphocytes [30], where comparable data are available. This must be due to the excellent degree of synchrony of green algae in a light/dark regime.…”

Section: Discussionmentioning

confidence: 99%

Deoxyribonucleotide Biosynthesis in Synchronous Algae Cells

Feller

Schimpff-Weiland

Follmann

1980

European Journal of Biochemistry

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Synchronous cells of the green alga, Scenedesmus obliquus, cultured in a 14‐h/10‐h light/dark regime, contain a peak of ribonucleoside‐diphosphate reductase activity and maximum deoxyribonucleoside 5′‐triphosphate concentrations at the 12th hour of the cell cycle, coinciding with DNA synthesis and preceding the formation of eight daughter cells. The intracellular dTTP pool reaches 4.5 pmol and the other pools 2–3 pmol/106 cells. Algal reductase activity is sensitive to cycloheximide, but not to lincomycin. These correlations demonstrate the functioning of the NDP → dNDP → dNTP pathway of DNA precursor biosynthesis in plant cells. In the presence of 20 μg 5‐fluorodeoxyuridine/ml, an inhibitor of thymidylate synthesis, the dTTP pool is rapidly depleted and DNA synthesis ceases. 5‐Fluorouracil and methotrexate produce similar effects. At the same time the ribonucleotide reductase activity and also the dATP pool are greatly increased, especially when fluorodeoxyuridine treatment is combined with continued illumination of the algae. In contrast, arabinosylcytosine, an inhibitor of DNA replication, has no effect on ribonucleotide reduction. The control of de novo enzyme synthesis in the eucaryotic algae therefore appears to depend on the presence of dTTP (or a related nucleotide), but not directly coupled to DNA synthesis. This interdependence resembles the situation observed in HeLa cells, while it may differ in detail from control mechanisms of ribonucleotide reductase studied in bacteria.

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“…The enzyme exhibits complex and strict allosteric control by a variety of positive and negative nucleotide effectors (9). In addition, the levels of RDR are tightly controlled so that greatly increased amounts of active enzyme are present at the onset of DNA synthesis, and high levels are maintained until DNA synthesis decreases (15,21,24).…”

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Chromosome-mediated gene transfer of hydroxyurea resistance and amplification of ribonucleotide reductase activity.

Lewis¹,

Srinivasan²

1983

Mol. Cell. Biol.

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Metaphase chromosomes purified from a hydroxyurea-resistant Chinese hamster cell line were able to transform recipient wild-type cells to hydroxyurea resistance at a frequency of 10-6. Approximately 60% of the resulting transformant clones gradually lost hydroxyurea resistance when cultivated for prolonged periods in the absence of drug. One transformant was subjected to serial selection in higher concentrations of hydroxyurea. The five cell lines generated exhibited increasing relative plating efficiency in the presence of the drug and a corresponding elevation in their cellular content of ribonucleotide reductase. The most resistant cell line had a 163-fold increase in relative plating efficiency and a 120-fold increase in enzyme activity when compared with the wild-type cell line. The highly hydroxyurea-resistant cell lines had strong electron paramagnetic resonance signals characteristic of an elevated level of the free radical present in the M2 subunit of ribonucleotide reductase. Two-dimensional electrophoresis of cellfree extracts from one of the resistant cell lines indicated that a 53,000-dalton protein was present in greatly elevated quantities when compared with the wildtype cell line. These data suggest that the hydroxyurea-resistant cell lines may contain an amplification of the gene for the M2 subunit of ribonucleotide reductase.

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Ribonucleotide reductase activity during the cell cycle of Saccharomyces cerevisiae

Cited by 68 publications

References 29 publications

Two genes differentially regulated in the cell cycle and by DNA-damaging agents encode alternative regulatory subunits of ribonucleotide reductase.

Two genes differentially regulated in the cell cycle and by DNA-damaging agents encode alternative regulatory subunits of ribonucleotide reductase.

Deoxyribonucleotide Biosynthesis in Synchronous Algae Cells

Chromosome-mediated gene transfer of hydroxyurea resistance and amplification of ribonucleotide reductase activity.

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