Three DNA polymerases, recruited by different mechanisms, carry out NER repair synthesis in human cells.
Nucleotide excision repair (NER) is the most versatile DNA repair system that deals with the major UV photoproducts in DNA, as well as many other DNA adducts. The early steps of NER are well understood, whereas the later steps of repair synthesis and ligation are not. In particular, which polymerases are definitely involved in repair synthesis and how they are recruited to the damaged sites has not yet been established. We report that, in human fibroblasts, approximately half of the repair synthesis requires both pol kappa and pol delta, and both polymerases can be recovered in the same repair complexes. Pol kappa is recruited to repair sites by ubiquitinated PCNA and XRCC1 and pol delta by the classical replication factor complex RFC1-RFC, together with a polymerase accessory factor, p66, and unmodified PCNA. The remaining repair synthesis is dependent on pol epsilon, recruitment of which is dependent on the alternative clamp loader CTF18-RFC.
Department of Molecular Medicine, Atomic Bomb Disease Institute, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4, Sakamoto, Nagasaki 852-8523, Japan. togi@nagasaki-u.ac.jp
Tomoo
T
Ogi
Siripan
S
Limsirichaikul
René M
RM
Overmeer
Marcel
M
Volker
Katsuya
K
Takenaka
Ross
R
Cloney
Yuka
Y
Nakazawa
Atsuko
A
Niimi
Yoshio
Y
Miki
Nicolaas G
NG
Jaspers
Leon H F
LH
Mullenders
Shunichi
S
Yamashita
Maria I
MI
Fousteri
Alan R
AR
Lehmann
D000074183:ATPases Associated with Diverse Cellular Activities; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D016922:Cellular Senescence; D004249:DNA Damage; D004257:DNA Polymerase II / Q000378:metabolism; D004258:DNA Polymerase III / Q000378:metabolism; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology* / Q000528:radiation effects; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism; D000075223:Poly-ADP-Ribose Binding Proteins; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D051818:Replication Protein C / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D044767:Ubiquitin-Protein Ligases; D054875:Ubiquitination; D014466:Ultraviolet Rays; D000076105:X-ray Repair Cross Complementing Protein 1; D000099270:Y-Family DNA Polymerases
D000074183:ATPases Associated with Diverse Cellular Activities; D002352:Carrier Proteins; D004257:DNA Polymerase II; D004258:DNA Polymerase III; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase; D009687:Nuclear Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D018809:Proliferating Cell Nuclear Antigen; D011993:Recombinant Fusion Proteins; D051818:Replication Protein C; D044767:Ubiquitin-Protein Ligases; D000076105:X-ray Repair Cross Complementing Protein 1; C477290:CHTF18 protein, human; C485954:RAD18 protein, human; C081632:RFC1 protein, human; C000619371:XRCC1 protein, human; C434055:POLD1 protein, human; C435720:POLD3 protein, human; C119525:POLE protein, human; C404302:POLK protein, human; D000099270:Y-Family DNA Polymerases