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J. Biol. Chem., Vol. 277, Issue 1, 746-754, January 4, 2002
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From the Department of Radiology and Cancer Biology, Nagasaki
University School of Dentistry, 1-7-1 Sakamoto,
Nagasaki 852-8588, Japan
Flap endonuclease 1 (FEN-1) is a 5'-3' flap
exo-/endonuclease that plays an important role in Okazaki fragment
maturation, nonhomologous end joining of double-stranded DNA breaks,
and long patch base excision repair. Here, we demonstrate that the wild type FEN-1 binds tightly to chromatin in conjunction with proliferating cell nuclear antigen (PCNA) recruitment after MMS treatment, and the
nuclease-defective FEN-1 increased the sensitivity of the cells to
methylmethane sulfonate (MMS) and to UV light but not to ionizing
radiation. In contrast, the cells expressing the nuclease-defective and
PCNA binding-defective double mutant FEN-1 exhibited sensitivities similar to those in the cells expressing the wild type FEN-1. MMS
treatment caused a prolonged delay of S phase progression and
impairment in colony-forming activity of cells expressing nuclease-defective FEN-1. A comet assay demonstrated that DNA repair
after MMS or UV treatment was impaired in the cells expressing nuclease-deficient FEN-1 but not in the cells with double-mutated FEN-1. Taken together, these findings suggest that FEN-1 plays an
essential role in the DNA repair processes in mammalian cells and
that this activity of FEN-1 is PCNA-dependent.
To whom correspondence should be addressed: Dept. of Radiology and
Cancer Biology Nagasaki University School of Dentistry 1-7-1 Sakamoto,
Nagasaki 852-8588, Japan. Tel.: 81 95 849 7707; Fax: 81 95 849 7711;
E-mail: taku@net.nagasaki-u.ac.jp.
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