Lcd1p recruits Mec1p to DNA lesions in vitro and in vivo

Mol Cell. 2002 Apr;9(4):857-69. doi: 10.1016/s1097-2765(02)00507-5.

Abstract

The Lcd1p/Mec1p complex is crucial for normal S phase progression and for signaling DNA damage. We show that Lcd1p/Ddc2p and Mec1p in cell extracts bind to DNA ends. Although Lcd1p binds DNA independently of Mec1p, recruitment of Mec1p to DNA requires Lcd1p. DNA binding by Lcd1p is also independent of Rad9p, Rad17p, and Rad24p. Recombinant Lcd1p binds DNA, and this is impaired by Lcd1p mutations that abrogate its in vivo functions. Furthermore, Mec1p is recruited to cdc13-induced DNA damage and HO endonuclease-induced double-strand breaks in vivo. This requires Lcd1p, and recruitment of Lcd1p/Mec1p to cdc13-induced damage is abolished by Lcd1p mutations that abrogate its in vivo functions. Recruitment of Lcd1p to these lesions is independent of Mec1p and Rad9p/Rad24p. Thus, recruitment of Mec1p to DNA lesions by Lcd1p is crucial for the DNA damage response.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Amino Acid Motifs
  • Cell Cycle Proteins / physiology
  • DNA Damage
  • DNA Repair / physiology*
  • DNA, Fungal / genetics*
  • DNA, Fungal / isolation & purification
  • DNA, Fungal / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Deoxyribonucleases, Type II Site-Specific / pharmacology
  • Evolution, Molecular
  • Fungal Proteins / genetics
  • Fungal Proteins / isolation & purification
  • Fungal Proteins / physiology*
  • In Vitro Techniques
  • Intracellular Signaling Peptides and Proteins
  • Macromolecular Substances
  • Microspheres
  • Nuclear Proteins
  • Phenotype
  • Phosphoproteins*
  • Phosphorylation
  • Protein Binding
  • Protein Kinases / genetics
  • Protein Kinases / physiology*
  • Protein Processing, Post-Translational
  • Protein-Serine-Threonine Kinases
  • Recombinant Fusion Proteins / physiology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / isolation & purification
  • Saccharomyces cerevisiae Proteins / metabolism
  • Saccharomyces cerevisiae Proteins / physiology*
  • Telomere-Binding Proteins*

Substances

  • Adaptor Proteins, Signal Transducing
  • Cdc13 protein, S cerevisiae
  • Cell Cycle Proteins
  • DNA, Fungal
  • DNA-Binding Proteins
  • Fungal Proteins
  • Intracellular Signaling Peptides and Proteins
  • LCD1 protein, S cerevisiae
  • Macromolecular Substances
  • Nuclear Proteins
  • Phosphoproteins
  • RAD17 protein, S cerevisiae
  • RAD24 protein, S cerevisiae
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • Telomere-Binding Proteins
  • rad9 protein
  • Protein Kinases
  • MEC1 protein, S cerevisiae
  • Protein-Serine-Threonine Kinases
  • HO protein, S cerevisiae
  • SCEI protein, S cerevisiae
  • Deoxyribonucleases, Type II Site-Specific