TY - JOUR

T1 - Incision of a 1,3-intrastrand d(GpTpG)-cisplatin adduct by nucleotide excision repair proteins from yeast

AU - Kong, Stephanie E.

AU - Svejstrup, Jesper Q.

N1 - 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.

PY - 2002/9/4

Y1 - 2002/9/4

N2 - 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).

AB - 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).

KW - DNA lesions

KW - Nucleotide excision repair

KW - Repairosome

KW - Saccharomyces cerevisiae

UR - http://www.scopus.com/inward/record.url?scp=0037019599&partnerID=8YFLogxK

U2 - 10.1016/S1568-7864(02)00080-0

DO - 10.1016/S1568-7864(02)00080-0

M3 - Journal article

C2 - 12509277

AN - SCOPUS:0037019599

VL - 1

SP - 731

EP - 741

JO - DNA Repair

JF - DNA Repair

SN - 1568-7864

IS - 9

ER -