Action of DNA repair endonuclease ERCC1/XPF in living cells

Science

Volumn 284, Issue 5416, 1999, Pages 958-961

  Houtsmuller, Adriaan B ^a   Rademakers, Suzanne ^a   Nigg, Alex L ^a   Hoogstraten, Deborah ^a   Hoeijmakers, Jan H J ^a   Vermeulen, Wim ^a  

^a ERASMUS MEDICAL CENTER   (Netherlands)

Author keywords

[No Author keywords available]

Indexed keywords

DNA; ENDONUCLEASE; GREEN FLUORESCENT PROTEIN;

ARTICLE; CONTROLLED STUDY; DNA REPAIR; ENZYME IMMOBILIZATION; EXCISION REPAIR; GENOME; HUMAN; HUMAN CELL; MOLECULAR SIZE; PRIORITY JOURNAL; PROTEIN ASSEMBLY; PROTEIN EXPRESSION; PROTEIN STABILITY;

ANIMALS; CELL LINE, TRANSFORMED; CELL NUCLEUS; CHO CELLS; CRICETINAE; DIFFUSION; DNA DAMAGE; DNA REPAIR; DNA-BINDING PROTEINS; ENDONUCLEASES; FLUORESCENCE; GREEN FLUORESCENT PROTEINS; HELA CELLS; HUMANS; LUMINESCENT PROTEINS; MICROSCOPY, CONFOCAL; MICROSCOPY, FLUORESCENCE; PROTEINS; RECOMBINANT FUSION PROTEINS; TRANSFECTION; ULTRAVIOLET RAYS;

CRICETINAE; CRICETULUS GRISEUS;

EID: 0033532342     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.284.5416.958     Document Type: Article

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50. Immunoblot analysis of total cell extracts (10 μg) and immunoprecipitation of cell lysates were performed as described (8). A standard colony-survival procedure for assessment of UV sensitivity (8) was applied to CHO-9 (parental repair-competent), 43-3B (ERCC1/repair-deficient), and 43-3B (ERCC1-GFP transfected) cells. At 5 days after UV irradiation (Philips TUV-lamp), the cell survival rate [(number of surviving treated colonies)/(number of surviving untreated colonies)] was plotted against UV dose.
51. We thank P. Berkhout for statistical advice; G. van Cappellen for technical advice; J. Essers and P. Doesburg for cell lines expressing, respectively, RAD52-GFP and AR-GFP; R. van Geloof for software; and R. van Driel, L. H. F. Mullenders, A. A. van Zeeland, and J. Trapman for helpful suggestions. Supported by the Dutch Cancer Society (KWF), the Dutch Scientific Organization (NWO) and by the Louis Jeantet Foundation.