p53-mediated DNA repair responses to UV radiation: studies of mouse cells lacking p53, p21, and/or gadd45 genes.
Human cells lacking functional p53 exhibit a partial deficiency in nucleotide excision repair (NER), the pathway for repair of UV-induced DNA damage. The global genomic repair (GGR) subpathway of NER, but not transcription-coupled repair (TCR), is mainly affected by p53 loss or inactivation. We have utilized mouse embryo fibroblasts (MEFs) lacking p53 genes or downstream effector genes of the p53 pathway, gadd45 (Gadd45a) or p21 (Cdkn1a), as well as MEFs lacking both gadd45 and p21 genes to address the potential contribution of these downstream effectors to p53-associated DNA repair. Loss of p53 or gadd45 had a pronounced effect on GGR, while p21 loss had only a marginal effect, determined by measurements of repair synthesis (unscheduled DNA synthesis), by immunoassays to detect removal of UV photoproducts from genomic DNA, and by assays determining strand-specific removal of CPDs from the mouse dhfr gene. Taken together, the evidence suggests a role for Gadd45, but relatively little role for p21, in DNA repair responses to UV radiation. Recent evidence suggests that Gadd45 binds to UV-damaged chromatin and may affect lesion accessibility. MEFs lacking p53 or gadd45 genes exhibited decreased colony-forming ability after UV radiation and cisplatin compared to wild-type MEFs, indicating their sensitivity to DNA damage. We provide evidence that Gadd45 affects chromatin remodelling of templates concurrent with DNA repair, thus indicating that Gadd45 may participate in the coupling between chromatin assembly and DNA repair.
Smith
M L
ML
Division of Basic Science, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Ford
J M
JM
Hollander
M C
MC
Bortnick
R A
RA
Amundson
S A
SA
Seo
Y R
YR
Deng
C X
CX
Hanawalt
P C
PC
Fornace
A J
AJ
D000818:Animals; D002843:Chromatin / Q000378:metabolism; D002945:Cisplatin / Q000494:pharmacology; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics*; D004260:DNA Repair / Q000235:genetics*; D004261:DNA Replication; D016158:Genes, p53; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D008817:Mice, Mutant Strains; D011506:Proteins / Q000235:genetics*; D011740:Pyrimidine Dimers / Q000378:metabolism; D016196:S Phase / Q000502:physiology; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014466:Ultraviolet Rays; D000097642:GADD45 Proteins
D002843:Chromatin; D002945:Cisplatin; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D047908:Intracellular Signaling Peptides and Proteins; D011506:Proteins; D011740:Pyrimidine Dimers; D013762:Tetrahydrofolate Dehydrogenase; D000097642:GADD45 Proteins; C493957:Cdkn1a protein, mouse; C058406:pyrimidine-pyrimidone dimer