Reduced extractability of the XPA DNA repair protein in ultraviolet light-irradiated mammalian cells.
The XPA protein is essential for both of the known modes of nucleotide excision repair (NER) in human cells: transcription-coupled repair (TCR) and global genome repair (GGR). In TCR, this protein is thought to be recruited to lesion sites in DNA at which RNA polymerase II is blocked and in GGR, by direct recognition of damages by repair protein complex containing XPC/HR23B or DNA damage-binding protein. However, details of the recruitment of the XPA protein in vivo are unknown. It was shown earlier that a portion of another NER protein, PCNA, which is completely extractable from non-S-phase mammalian nuclei, becomes insoluble after ultraviolet (UV) light irradiation and cannot be extracted by methanol or buffer containing Triton X-100. In the present study, we have found that UV light irradiation of human or Chinese hamster cells leads to decrease of extractability of the XPA protein by Triton X-100. Maximal insolubilization of the XPA protein is observed 1-4 h after irradiation but it is not detectable by 22 h. This effect is dose-dependent for UV light from 2.5 to 15 J/m(2) and is unaffected by the pre-treatment of cells with sodium butyrate, an inhibitor of histone deacetylation. The UV light-induced insolubilization of the XPA protein was also observed in two lines of Cockayne syndrome complementation group A cells, indicating that the effect is not dependent upon TCR. The results are discussed in relation to possible mechanisms of NER.
Svetlova
M
M
Institute of Cytology of the Russian Academy of Sciences, 194064, St. Petersburg, Russia.
Nikiforov
A
A
Solovjeva
L
L
Pleskach
N
N
Tomilin
N
N
Hanawalt
P C
PC
D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D003057:Cockayne Syndrome / Q000235:genetics / Q000378:metabolism; D006224:Cricetinae; D004249:DNA Damage; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000528:radiation effects*; D003902:Detergents / Q000494:pharmacology; D004307:Dose-Response Relationship, Radiation; D005347:Fibroblasts / Q000737:chemistry / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D017830:Octoxynol / Q000494:pharmacology; D012995:Solubility / Q000187:drug effects / Q000528:radiation effects; D013997:Time Factors; D014466:Ultraviolet Rays; D051760:Xeroderma Pigmentosum Group A Protein
D004268:DNA-Binding Proteins; D003902:Detergents; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein; D017830:Octoxynol