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Science 7 May 1999:
Vol. 284. no. 5416, pp. 958 - 961
DOI: 10.1126/science.284.5416.958
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Reports

Action of DNA Repair Endonuclease ERCC1/XPF in Living Cells

Adriaan B. Houtsmuller, 1* Suzanne Rademakers, 2 Alex L. Nigg, 1 Deborah Hoogstraten, 2 Jan H. J. Hoeijmakers, 2 Wim Vermeulen 2*

To study the nuclear organization and dynamics of nucleotide excision repair (NER), the endonuclease ERCC1/XPF (for excision repair cross complementation group 1/xeroderma pigmentosum group F) was tagged with green fluorescent protein and its mobility was monitored in living Chinese hamster ovary cells. In the absence of DNA damage, the complex moved freely through the nucleus, with a diffusion coefficient (15 ± 5 square micrometers per second) consistent with its molecular size. Ultraviolet light-induced DNA damage caused a transient dose-dependent immobilization of ERCC1/XPF, likely due to engagement of the complex in a single repair event. After 4 minutes, the complex regained mobility. These results suggest (i) that NER operates by assembly of individual NER factors at sites of DNA damage rather than by preassembly of holocomplexes and (ii) that ERCC1/XPF participates in repair of DNA damage in a distributive fashion rather than by processive scanning of large genome segments.

1 Department of Pathology (Josephine Nefkens Institute) and
2 Department of Cell Biology and Genetics (Medical Genetics Center, CBG), Erasmus University, Post Office Box 1738, 3000 DR Rotterdam, Netherlands.
*   These authors contributed equally to this work.

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