Endogenous DNA abasic sites cause cell death in the absence of Apn1, Apn2 and Rad1/Rad10 in Saccharomyces cerevisiae.
In Saccharomyces cerevisiae, mutations in APN1, APN2 and either RAD1 or RAD10 genes are synthetic lethal. In fact, apn1 apn2 rad1 triple mutants can form microcolonies of approximately 300 cells. Expression of Nfo, the bacterial homologue of Apn1, suppresses the lethality. Turning off the expression of Nfo induces G(2)/M cell cycle arrest in an apn1 apn2 rad1 triple mutant. The activation of this checkpoint is RAD9 dependent and allows residual DNA repair. The Mus81/Mms4 complex was identified as one of these back-up repair activities. Furthermore, inactivation of Ntg1, Ntg2 and Ogg1 DNA N-glycosylase/AP lyases in the apn1 apn2 rad1 background delayed lethality, allowing the formation of minicolonies of approximately 10(5) cells. These results demonstrate that, under physiological conditions, endogenous DNA damage causes death in cells deficient in Apn1, Apn2 and Rad1/Rad10 proteins. We propose a model in which endogenous DNA abasic sites are converted into 3'-blocked single-strand breaks (SSBs) by DNA N-glycosylases/AP lyases. Therefore, we suggest that the essential and overlapping function of Apn1, Apn2, Rad1/Rad10 and Mus81/Mms4 is to repair 3'-blocked SSBs using their 3'-phosphodiesterase activity or their 3'-flap endonuclease activity, respectively.
Guillet
Marie
M
CEA, DSV, Département de Radiobiologie et Radiopathologie, UMR217 CNRS Radiobiologie Moléculaire et Cellulaire, BP6, F-92265 Fontenay aux Roses, France.
Boiteux
Serge
S
D000626:Aminopeptidases / Q000235:genetics*; D016923:Cell Death; D002469:Cell Separation; D004247:DNA / Q000737:chemistry*; D004249:DNA Damage; D045643:DNA Repair Enzymes; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D004268:DNA-Binding Proteins; D043382:DNA-Formamidopyrimidine Glycosylase; D004305:Dose-Response Relationship, Drug; D004720:Endonucleases / Q000235:genetics*; D005434:Flow Cytometry; D005656:Fungal Proteins / Q000235:genetics*; D016195:G2 Phase; D019476:Insect Proteins / Q000235:genetics*; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008938:Mitosis; D009153:Mutagens / Q000494:pharmacology; D009154:Mutation; D009699:N-Glycosyl Hydrolases / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D015719:Single-Strand Specific DNA and RNA Endonucleases; D013997:Time Factors
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D019476:Insect Proteins; D009153:Mutagens; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; D008741:Methyl Methanesulfonate; D004720:Endonucleases; C069266:RAD1 protein, S cerevisiae; C064162:RAD10 protein, S cerevisiae; D015719:Single-Strand Specific DNA and RNA Endonucleases; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase; D000626:Aminopeptidases; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; C101825:NTG1 protein, S cerevisiae; C116827:NTG2 protein, S cerevisiae; D045643:DNA Repair Enzymes