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PMID: 9670034 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mec1p is essential for phosphorylation of the yeast DNA damage checkpoint protein Ddc1p, which physically interacts with Mec3p.

The EMBO journal ·Vol. 17 ·No. 14 ·1998-07-15 ·Pages 4199-209

Paciotti V, Lucchini G, Plevani P, Longhese MP

Abstract

Checkpoints prevent DNA replication or nuclear division when chromosomes are damaged. The Saccharomyces cerevisiae DDC1 gene belongs to the RAD17, MEC3 and RAD24 epistasis group which, together with RAD9, is proposed to act at the beginning of the DNA damage checkpoint pathway. Ddc1p is periodically phosphorylated during unperturbed cell cycle and hyperphosphorylated in response to DNA damage. We demonstrate that Ddc1p interacts physically in vivo with Mec3p, and this interaction requires Rad17p. We also show that phosphorylation of Ddc1p depends on the key checkpoint protein Mec1p and also on Rad24p, Rad17p and Mec3p. This suggests that Mec1p might act together with the Rad24 group of proteins at an early step of the DNA damage checkpoint response. On the other hand, Ddc1p phosphorylation is independent of Rad53p and Rad9p. Moreover, while Ddc1p is required for Rad53p phosphorylation, it does not play any major role in the phosphorylation of the anaphase inhibitor Pds1p, which requires RAD9 and MEC1. We suggest that Rad9p and Ddc1p might function in separated branches of the DNA damage checkpoint pathway, playing different roles in determining Mec1p activity and/or substrate specificity.

MeSH Terms
Cell Cycle/physiology Cell Cycle Proteins/genetics,metabolism DNA Damage/physiology DNA, Fungal/genetics Fungal Proteins/genetics,metabolism Intracellular Signaling Peptides and Proteins Nuclear Proteins/metabolism Phosphorylation Protein Serine-Threonine Kinases Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Securin
Chemicals
Cell Cycle Proteins DNA, Fungal Fungal Proteins Intracellular Signaling Peptides and Proteins MEC3 protein, S cerevisiae Nuclear Proteins PDS1 protein, S cerevisiae Saccharomyces cerevisiae Proteins Securin MEC1 protein, S cerevisiae Protein Serine-Threonine Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Paciotti V
Dipartimento di Genetica e di Biologia dei Microrganismi, Università degli Studi di Milano, Via Celoria 26, 20133 Milano, Italy.
Lucchini G
Plevani P
Longhese M P
References (51)
51 references, click to expand
  1. Cell cycle checkpoints: preventing an identity crisis.
    Science. 1996 Dec 6;274(5293):1664-72 PMID: 8939848
  2. Spk1/Rad53 is regulated by Mec1-dependent protein phosphorylation in DNA replication and damage checkpoint pathways.
    Genes Dev. 1996 Feb 15;10(4):395-406 PMID: 8600024
  3. The cellular responses to DNA damage.
    Trends Cell Biol. 1995 Jan;5(1):32-40 PMID: 14731431
  4. RAD9, RAD17, and RAD24 are required for S phase regulation in Saccharomyces cerevisiae in response to DNA damage.
    Genetics. 1997 Jan;145(1):45-62 PMID: 9017389
  5. The B-type cyclin kinase inhibitor p40SIC1 controls the G1 to S transition in S. cerevisiae.
    Cell. 1994 Oct 21;79(2):233-44 PMID: 7954792
  6. Menage à trois: double strand break repair, V(D)J recombination and DNA-PK.
    Bioessays. 1995 Nov;17(11):949-57 PMID: 8526889
  7. The Schizosaccharomyces pombe rad3 checkpoint gene.
    EMBO J. 1996 Dec 2;15(23):6641-51 PMID: 8978690
  8. DUN1 encodes a protein kinase that controls the DNA damage response in yeast.
    Cell. 1993 Dec 17;75(6):1119-27 PMID: 8261511
  9. A single ataxia telangiectasia gene with a product similar to PI-3 kinase.
    Science. 1995 Jun 23;268(5218):1749-53 PMID: 7792600
  10. Anaphase initiation in Saccharomyces cerevisiae is controlled by the APC-dependent degradation of the anaphase inhibitor Pds1p.
    Genes Dev. 1996 Dec 15;10(24):3081-93 PMID: 8985178
  11. Molecular cloning and analysis of Schizosaccharomyces pombe rad9, a gene involved in DNA repair and mutagenesis.
    Mol Gen Genet. 1992 Apr;232(3):367-76 PMID: 1588907
  12. DNA polymerase epsilon links the DNA replication machinery to the S phase checkpoint.
    Cell. 1995 Jan 13;80(1):29-39 PMID: 7813016
  13. RAD9-dependent G1 arrest defines a second checkpoint for damaged DNA in the cell cycle of Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):7985-9 PMID: 8367452
  14. Rad53-dependent phosphorylation of Swi6 and down-regulation of CLN1 and CLN2 transcription occur in response to DNA damage in Saccharomyces cerevisiae.
    Genes Dev. 1997 Nov 15;11(22):3032-45 PMID: 9367985
  15. SPK1 is an essential S-phase-specific gene of Saccharomyces cerevisiae that encodes a nuclear serine/threonine/tyrosine kinase.
    Mol Cell Biol. 1993 Sep;13(9):5829-42 PMID: 8355715
  16. Hus1p, a conserved fission yeast checkpoint protein, interacts with Rad1p and is phosphorylated in response to DNA damage.
    EMBO J. 1998 Apr 1;17(7):2055-66 PMID: 9524127
  17. Complete transposition requires four active monomers in the mu transposase tetramer.
    Genes Dev. 1994 Oct 15;8(20):2416-28 PMID: 7958906
  18. Cell cycle control and cancer.
    Science. 1994 Dec 16;266(5192):1821-8 PMID: 7997877
  19. DNA repair mutants defining G2 checkpoint pathways in Schizosaccharomyces pombe.
    EMBO J. 1992 Apr;11(4):1343-50 PMID: 1563350
  20. Mitotic checkpoint genes in budding yeast and the dependence of mitosis on DNA replication and repair.
    Genes Dev. 1994 Mar 15;8(6):652-65 PMID: 7926756
  21. Single-stranded DNA arising at telomeres in cdc13 mutants may constitute a specific signal for the RAD9 checkpoint.
    Mol Cell Biol. 1995 Nov;15(11):6128-38 PMID: 7565765
  22. Characterization of G1 checkpoint control in the yeast Saccharomyces cerevisiae following exposure to DNA-damaging agents.
    Genetics. 1994 Oct;138(2):271-81 PMID: 7828811
  23. Yeast pip3/mec3 mutants fail to delay entry into S phase and to slow DNA replication in response to DNA damage, and they define a functional link between Mec3 and DNA primase.
    Mol Cell Biol. 1996 Jul;16(7):3235-44 PMID: 8668138
  24. Distinct roles of yeast MEC and RAD checkpoint genes in transcriptional induction after DNA damage and implications for function.
    Mol Biol Cell. 1996 May;7(5):703-18 PMID: 8744945
  25. Dpb11, which interacts with DNA polymerase II(epsilon) in Saccharomyces cerevisiae, has a dual role in S-phase progression and at a cell cycle checkpoint.
    Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11791-5 PMID: 8524850
  26. The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae.
    Science. 1988 Jul 15;241(4863):317-22 PMID: 3291120
  27. Yeast checkpoint genes in DNA damage processing: implications for repair and arrest.
    Science. 1995 Dec 1;270(5241):1488-91 PMID: 7491494
  28. Cell cycle arrest of cdc mutants and specificity of the RAD9 checkpoint.
    Genetics. 1993 May;134(1):63-80 PMID: 8514150
  29. Rfc5, a small subunit of replication factor C complex, couples DNA replication and mitosis in budding yeast.
    Proc Natl Acad Sci U S A. 1996 Jul 9;93(14):7048-52 PMID: 8692942
  30. Pds1p, an inhibitor of anaphase in budding yeast, plays a critical role in the APC and checkpoint pathway(s).
    J Cell Biol. 1996 Apr;133(1):99-110 PMID: 8601617
  31. A checkpoint regulates the rate of progression through S phase in S. cerevisiae in response to DNA damage.
    Cell. 1995 Sep 8;82(5):841-7 PMID: 7671311
  32. Cloning and characterisation of the rad9 DNA repair gene from Schizosaccharomyces pombe.
    Nucleic Acids Res. 1991 Jul 11;19(13):3525-31 PMID: 1852603
  33. RAD9 and DNA polymerase epsilon form parallel sensory branches for transducing the DNA damage checkpoint signal in Saccharomyces cerevisiae.
    Genes Dev. 1996 Oct 15;10(20):2632-43 PMID: 8895664
  34. The novel DNA damage checkpoint protein ddc1p is phosphorylated periodically during the cell cycle and in response to DNA damage in budding yeast.
    EMBO J. 1997 Sep 1;16(17):5216-26 PMID: 9311982
  35. A role for DNA primase in coupling DNA replication to DNA damage response.
    EMBO J. 1997 Feb 3;16(3):639-50 PMID: 9034345
  36. The anaphase inhibitor of Saccharomyces cerevisiae Pds1p is a target of the DNA damage checkpoint pathway.
    Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14361-6 PMID: 9405617
  37. Rfc5, a replication factor C component, is required for regulation of Rad53 protein kinase in the yeast checkpoint pathway.
    Mol Cell Biol. 1997 Oct;17(10):5905-14 PMID: 9315648
  38. A human homolog of the Schizosaccharomyces pombe rad9+ checkpoint control gene.
    Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13890-5 PMID: 8943031
  39. The Polo-like kinase Cdc5p and the WD-repeat protein Cdc20p/fizzy are regulators and substrates of the anaphase promoting complex in Saccharomyces cerevisiae.
    EMBO J. 1998 Mar 2;17(5):1336-49 PMID: 9482731
  40. cDNA cloning and gene mapping of a candidate human cell cycle checkpoint protein.
    Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2850-5 PMID: 8610130
  41. When checkpoints fail.
    Cell. 1997 Feb 7;88(3):315-21 PMID: 9039258
  42. Regulation of RAD53 by the ATM-like kinases MEC1 and TEL1 in yeast cell cycle checkpoint pathways.
    Science. 1996 Jan 19;271(5247):357-60 PMID: 8553072
  43. Interaction of DNA-dependent protein kinase with DNA and with Ku: biochemical and atomic-force microscopy studies.
    EMBO J. 1997 Aug 15;16(16):5098-112 PMID: 9305651
  44. Control of cell cycle arrest by the Mec1sc/Rad3sp DNA structure checkpoint pathway.
    Curr Opin Genet Dev. 1997 Feb;7(1):93-8 PMID: 9024628
  45. A novel role for the budding yeast RAD9 checkpoint gene in DNA damage-dependent transcription.
    EMBO J. 1996 Aug 1;15(15):3912-22 PMID: 8670896
  46. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites.
    Gene. 1988 Dec 30;74(2):527-34 PMID: 3073106
  47. DNA double-strand breaks caused by replication arrest.
    EMBO J. 1997 Jan 15;16(2):430-8 PMID: 9029161
  48. From DNA damage to cell cycle arrest and suicide: a budding yeast perspective.
    Curr Opin Genet Dev. 1996 Feb;6(1):4-11 PMID: 8791492
  49. The Saccharomyces cerevisiae MEC1 gene, which encodes a homolog of the human ATM gene product, is required for G1 arrest following radiation treatment.
    J Bacteriol. 1996 Oct;178(19):5841-3 PMID: 8824640
  50. SOS-regulated proteins in translesion DNA synthesis and mutagenesis.
    Trends Biochem Sci. 1995 Oct;20(10):416-20 PMID: 8533155
  51. The 70 kDa subunit of replication protein A is required for the G1/S and intra-S DNA damage checkpoints in budding yeast.
    Nucleic Acids Res. 1996 Sep 15;24(18):3533-7 PMID: 8836179
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.14.4199
Published
1998-07-15
Pages
4199-209
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170752
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/17.14.4199 PMC Full Text View journal details
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