A novel role in DNA metabolism for the binding of Fen1/Rad27 to PCNA and implications for genetic risk.
Fen1/Rad27 nuclease activity, which is important in DNA metabolism, is stimulated by proliferating cell nuclear antigen (PCNA) in vitro. The in vivo role of the PCNA interaction was investigated in the yeast Rad27. A nuclease-defective rad27 mutation had a dominant-negative effect that was suppressed by a mutation in the PCNA binding site, thereby demonstrating the importance of the Rad27-PCNA interaction. The PCNA-binding defect alone had little effect on mutation, recombination, and the methyl methanesulfonate (MMS) response in repair-competent cells, but it greatly amplified the MMS sensitivity of a rad51 mutant. Furthermore, the PCNA binding mutation resulted in lethality when combined with a homozygous or even a heterozygous pol3-01 mutation in the 3'-->5' exonuclease domain of DNA polymerase delta. These results suggest that phenotypically mild polymorphisms in DNA metabolic proteins can have dramatic consequences when combined.
Gary
R
R
Life Sciences Division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545, USA.
Park
M S
MS
Nolan
J P
JP
Cornelius
H L
HL
Kozyreva
O G
OG
Tran
H T
HT
Lobachev
K S
KS
Resnick
M A
MA
Gordenin
D A
DA
D001665:Binding Sites; D000071877:Checkpoint Kinase 1; D004249:DNA Damage; D004258:DNA Polymerase III / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair; D004261:DNA Replication; D004271:DNA, Fungal / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D004351:Drug Resistance / Q000235:genetics; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology*; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008957:Models, Genetic; D009153:Mutagens / Q000494:pharmacology; D011110:Polymorphism, Genetic; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D011485:Protein Binding; D011494:Protein Kinases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D051135:Rad51 Recombinase; D012306:Risk; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D009153:Mutagens; D018809:Proliferating Cell Nuclear Antigen; D029701:Saccharomyces cerevisiae Proteins; D008741:Methyl Methanesulfonate; D011494:Protein Kinases; D000071877:Checkpoint Kinase 1; C495151:RAD51 protein, S cerevisiae; D051135:Rad51 Recombinase; D004258:DNA Polymerase III; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V