DNA Damage in the Nucleosome Core Is Refractory to Repair by Human Excision Nuclease

Abstract

To investigate the effect of nucleosomes on nucleotide excision repair in humans, we prepared a mononucleosome containing a (6-4) photoproduct in the nucleosome core and examined its repair with the reconstituted human excision nuclease system and with cell extracts. Nucleosomal DNA is repaired at a rate of about 10% of that for naked DNA in both systems. These results are in agreement with in vivo data showing a considerably slower rate of repair of overall genomic DNA relative to that for transcriptionally active DNA. Furthermore, our results indicate that the first-order packing of DNA in nucleosomes is a primary determinant of slow repair of DNA in chromatin.

ACKNOWLEDGMENTS

We thank T. Bessho, L. Lindsey-Boltz, J. Reardon, and C. Selby for useful discussions and J. Reardon and C. Selby for critical reading of the manuscript.

This work was supported by National Institutes of Health grant GM 32833.