Novel functional requirements for non-homologous DNA end joining in Schizosaccharomyces pombe
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In: EMBO Journal, Vol. 20, No. 1-2, 15.01.2001, p. 210-21.
Research output: Contribution to journal › Article › peer-review
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T1 - Novel functional requirements for non-homologous DNA end joining in Schizosaccharomyces pombe
AU - Manolis, K G
AU - Nimmo, E R
AU - Hartsuiker, E
AU - Carr, A M
AU - Jeggo, P A
AU - Allshire, R C
PY - 2001/1/15
Y1 - 2001/1/15
N2 - 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.
AB - 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.
KW - Animals
KW - Antigens, Nuclear
KW - Base Sequence
KW - Bleomycin
KW - Cell Nucleus
KW - DNA Damage
KW - DNA Helicases
KW - DNA Ligase ATP
KW - DNA Ligases
KW - DNA Repair
KW - DNA, Fungal
KW - DNA-Binding Proteins
KW - Fungal Proteins
KW - Gamma Rays
KW - Gene Silencing
KW - Ku Autoantigen
KW - Mammals
KW - Molecular Sequence Data
KW - Nuclear Proteins
KW - Restriction Mapping
KW - Saccharomyces cerevisiae Proteins
KW - Schizosaccharomyces
KW - Telomere
KW - Temperature
KW - Transcription Factors
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1093/emboj/20.1.210
DO - 10.1093/emboj/20.1.210
M3 - Article
C2 - 11226171
VL - 20
SP - 210
EP - 221
JO - EMBO Journal
JF - EMBO Journal
SN - 0261-4189
IS - 1-2
ER -