Novel functional requirements for non-homologous DNA end joining in Schizosaccharomyces pombe.
DNA double strand break (DSB) repair by non-homologous end joining (NHEJ) in mammalian cells requires the Ku70-Ku80 heterodimer, the DNA-PK catalytic subunit DNA-PKcs, as well as DNA ligase IV and Xrcc4. NHEJ of plasmid DSBs in Saccharomyces cerevisiae requires Ku, Xrcc4 and DNA ligase IV, as well as Mre11, Rad50, Xrs2 and DNA damage checkpoint proteins. Saccharomyces cerevisiae Ku is also required for telomere length maintenance and transcriptional silencing. We have characterized NHEJ in Schizosaccharomyces pombe using an extrachromosomal assay and find that, as anticipated, it is Ku70 and DNA ligase IV dependent. Unexpectedly, we find that Rad32, Rad50 (the S.pombe homologues of Mre11 and Rad50, respectively) and checkpoint proteins are not required for NHEJ. Furthermore, although S.pombe Ku70 is required for maintenance of telomere length, it is dispensable for transcriptional silencing at telomeres and is located throughout the nucleus rather than concentrated at the telomeres. Together, these results provide insight into the mechanism of NHEJ and contrast significantly with recent studies in S.cerevisiae.
Manolis
K G
KG
MRC Cell Mutation Unit, University of Sussex, Falmer, Sussex BN1, UK.
Nimmo
E R
ER
Hartsuiker
E
E
Carr
A M
AM
Jeggo
P A
PA
Allshire
R C
RC
D000818:Animals; D034961:Antigens, Nuclear; D001483:Base Sequence; D001761:Bleomycin / Q000494:pharmacology; D002467:Cell Nucleus / Q000235:genetics / Q000502:physiology; D004249:DNA Damage; D004265:DNA Helicases; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000378:metabolism*; D004260:DNA Repair; D004271:DNA, Fungal / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D005720:Gamma Rays; D020868:Gene Silencing; D000072200:Ku Autoantigen; D008322:Mammals; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D015183:Restriction Mapping; D029701:Saccharomyces cerevisiae Proteins; D012568:Schizosaccharomyces / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D016615:Telomere / Q000235:genetics / Q000502:physiology; D013696:Temperature; D014157:Transcription Factors / Q000378:metabolism
D034961:Antigens, Nuclear; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C000607891:DNL4 protein, S cerevisiae; D005656:Fungal Proteins; C000607893:LIG4 protein, human; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C081647:high affinity DNA-binding factor, S cerevisiae; D001761:Bleomycin; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen; D011088:DNA Ligases; D000072481:DNA Ligase ATP