Incision of a 1,3-intrastrand d(GpTpG)-cisplatin adduct by nucleotide excision repair proteins from yeast
Research output: Contribution to journal › Journal article › Research › peer-review
The protein machinery responsible for nucleotide excision repair (NER) is highly conserved from yeast to man. NER can be reconstituted with purified proteins, and the incision sites around a defined DNA lesion have been defined to the nucleotide level in a mammalian NER system. Here, we reconstitute NER in yeast whole cell extracts, as well as with partially purified yeast NER components. We show that NER activity can be isolated partly as a large protein complex, and map the sites of nucleotide incision around a cisplatin-induced DNA lesion. Our data indicate that yeast NER proteins excise an oligonucleotide of 23-26 bases containing the DNA lesion (rather than 26-30 bases as in humans), and that the 3′ incision occurs around position 17 (rather than at position 9 as in humans).
| Original language | English |
|---|---|
| Journal | DNA Repair |
| Volume | 1 |
| Issue number | 9 |
| Pages (from-to) | 731-741 |
| Number of pages | 11 |
| ISSN | 1568-7864 |
| DOIs | |
| Publication status | Published - 4 Sep 2002 |
| Externally published | Yes |
Bibliographical note
Funding Information:
This work was supported by an Overseas Research Grant, Bath Fellowship to S.E.K. and a grant from Cancer Research UK to J.Q.S. We thank Rick Wood and members of his laboratory for advice, reagents and enthusiastic encouragement when they were at Clare Hall Laboratories. Mahmud Shivji and Rick Wood’s work on a direct end-labelling system to study NER in human cell extracts was instrumental in making it possible to obtain the described system in yeast. We also acknowledge the service departments, especially the Fermentation Service at Cancer Research UK for their helpful assistance. Cancer Research UK London Research Institute comprises the Lincoln’s Inn Fields and Clare Hall Laboratories of the former Imperial Cancer Research Fund following the merger of the ICRF with the Cancer Research Campaign in February 2002.
- DNA lesions, Nucleotide excision repair, Repairosome, Saccharomyces cerevisiae
Research areas
ID: 331042274