1. Mck1 defines a key S-phase checkpoint effector in response to various degrees of replication threats
- Author
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Jingjing Zhang, Xiaojing Liu, Qinhong Cao, Li-Lin Du, Yanling Niu, Beidong Liu, Zhen Li, Jiaxin Zhang, Wenya Hou, Huiqiang Lou, Xuejiao Jin, Jiani Li, Sushma Sharma, and Xiaoli Li
- Subjects
Cancer Research ,Cell- och molekylärbiologi ,Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) ,Synthesis Phase ,Gene Expression ,Cell Cycle Proteins ,Yeast and Fungal Models ,QH426-470 ,Biochemistry ,Gene Knockout Techniques ,Glycogen Synthase Kinase 3 ,0302 clinical medicine ,Transcription (biology) ,Gene Expression Regulation, Fungal ,Transcriptional regulation ,Hydroxyurea ,Cell Cycle and Cell Division ,Post-Translational Modification ,Phosphorylation ,Promoter Regions, Genetic ,Genetics (clinical) ,Cellular Stress Responses ,Regulation of gene expression ,0303 health sciences ,Nucleotides ,Effector ,Transcriptional Control ,Messenger RNA ,Eukaryota ,In Vitro Kinase Assay ,Cell biology ,Nucleic acids ,Ribonucleotide reductase ,Bioassays and Physiological Analysis ,Experimental Organism Systems ,Cell Processes ,S Phase Cell Cycle Checkpoints ,Perspective ,Saccharomyces Cerevisiae ,Research Article ,DNA Replication ,Saccharomyces cerevisiae Proteins ,DNA damage ,Repressor ,Protein Serine-Threonine Kinases ,Biology ,Research and Analysis Methods ,03 medical and health sciences ,Saccharomyces ,Model Organisms ,Ribonucleotide Reductases ,Genetics ,Gene Regulation ,Molecular Biology ,Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci) ,Ecology, Evolution, Behavior and Systematics ,030304 developmental biology ,Enzyme Assays ,Biology and life sciences ,Organisms ,Fungi ,Proteins ,Promoter ,DNA ,Cell Biology ,Ribonucleotides ,Yeast ,Repressor Proteins ,Animal Studies ,RNA ,Biochemical Analysis ,030217 neurology & neurosurgery ,Cell and Molecular Biology - Abstract
The S-phase checkpoint plays an essential role in regulation of the ribonucleotide reductase (RNR) activity to maintain the dNTP pools. How eukaryotic cells respond appropriately to different levels of replication threats remains elusive. Here, we have identified that a conserved GSK-3 kinase Mck1 cooperates with Dun1 in regulating this process. Deleting MCK1 sensitizes dun1Δ to hydroxyurea (HU) reminiscent of mec1Δ or rad53Δ. While Mck1 is downstream of Rad53, it does not participate in the post-translational regulation of RNR as Dun1 does. Mck1 phosphorylates and releases the Crt1 repressor from the promoters of DNA damage-inducible genes as RNR2-4 and HUG1. Hug1, an Rnr2 inhibitor normally silenced, is induced as a counterweight to excessive RNR. When cells suffer a more severe threat, Mck1 inhibits HUG1 transcription. Consistently, only a combined deletion of HUG1 and CRT1, confers a dramatic boost of dNTP levels and the survival of mck1Δdun1Δ or mec1Δ cells assaulted by a lethal dose of HU. These findings reveal the division-of-labor between Mck1 and Dun1 at the S-phase checkpoint pathway to fine-tune dNTP homeostasis., Author summary The appropriate amount and balance of four dNTPs are crucial for all cells correctly copying and passing on their genetic material generation by generation. Eukaryotes have developed an alert and response system to deal with the disturbance. Here, we uncovered a second-level effector branch. It is activated by the upstream surveillance kinase cascade, which can induce the expression of dNTP-producing enzymes. It can also reduce the inhibitor of these enzymes to further boost their activity according to the degrees of threats. These findings suggest a multi-level response system to guarantee the appropriate dNTP supply, which is essential to maintain genetic stability under various environmental challenges.
- Published
- 2019