Purification and cloning of a nucleotide excision repair complex involving the xeroderma pigmentosum group C protein and a human homologue of yeast RAD23.
Complementation group C of xeroderma pigmentosum (XP) represents one of the most common forms of this cancer-prone DNA repair syndrome. The primary defect is located in the subpathway of the nucleotide excision repair system, dealing with the removal of lesions from the non-transcribing sequences ('genome-overall' repair). Here we report the purification to homogeneity and subsequent cDNA cloning of a repair complex by in vitro complementation of the XP-C defect in a cell-free repair system containing UV-damaged SV40 minichromosomes. The complex has a high affinity for ssDNA and consists of two tightly associated proteins of 125 and 58 kDa. The 125 kDa subunit is an N-terminally extended version of previously reported XPCC gene product which is thought to represent the human homologue of the Saccharomyces cerevisiae repair gene RAD4. The 58 kDa species turned out to be a human homologue of yeast RAD23. Unexpectedly, a second human counterpart of RAD23 was identified. All RAD23 derivatives share a ubiquitin-like N-terminus. The nature of the XP-C defect implies that the complex exerts a unique function in the genome-overall repair pathway which is important for prevention of skin cancer.
Masutani
C
C
Cellular Physiology Laboratory, Institute of Physical and Chemical Research (RIKEN), Saitama, Japan.
Sugasawa
K
K
Yanagisawa
J
J
Sonoyama
T
T
Ui
M
M
Enomoto
T
T
Takio
K
K
Tanaka
K
K
van der Spek
P J
PJ
Bootsma
D
D
D000595:Amino Acid Sequence; D001483:Base Sequence; D004249:DNA Damage; D004260:DNA Repair; D045643:DNA Repair Enzymes; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D005656:Fungal Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D029702:Schizosaccharomyces pombe Proteins; D017386:Sequence Homology, Amino Acid; D013539:Simian virus 40 / Q000235:genetics; D011503:Transglutaminases; D014452:Ubiquitins / Q000235:genetics; D014466:Ultraviolet Rays
D018076:DNA, Complementary; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C082463:RAD4 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D014452:Ubiquitins; C086985:RAD23A protein, human; C086984:XPC protein, human; D011503:Transglutaminases; D045643:DNA Repair Enzymes