PCNA mono-ubiquitination and activation of translesion DNA polymerases by DNA polymerase {alpha}.
Translesion DNA synthesis (TLS) involves PCNA mono-ubiquitination and TLS DNA polymerases (pols). Recent evidence has shown that the mono-ubiquitination is induced not only by DNA damage but also by other factors that induce stalling of the DNA replication fork. We studied the effect of spontaneous DNA replication errors on PCNA mono-ubiquitination and TLS induction. In the pol1L868F strain, which expressed an error-prone pol alpha, PCNA was spontaneously mono-ubiquitinated. Pol alpha L868F had a rate-limiting step at the extension from mismatched primer termini. Electron microscopic observation showed the accumulation of a single-stranded region at the DNA replication fork in yeast cells. For pol alpha errors, pol zeta participated in a generation of +1 frameshifts. Furthermore, in the pol1L868F strain, UV-induced mutations were lower than in the wild-type and a pol delta mutant strain (pol3-5DV), and deletion of the RAD30 gene (pol eta) suppressed this defect. These data suggest that nucleotide misincorporation by pol alpha induces exposure of single-stranded DNA, PCNA mono-ubiquitination and activates TLS pols.
Suzuki
Motoshi
M
Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine, Japan. msuzuki@med.nagoya-u.ac.jp
Niimi
Atsuko
A
Limsirichaikul
Siripan
S
Tomida
Shuta
S
Miao Huang
Qin
Q
Izuta
Shunji
S
Usukura
Jiro
J
Itoh
Yasutomo
Y
Hishida
Takashi
T
Akashi
Tomohiro
T
Nakagawa
Yoshiyuki
Y
Kikuchi
Akihiko
A
Pavlov
Youri
Y
Murate
Takashi
T
Takahashi
Takashi
T
D020137:Base Pair Mismatch; D002470:Cell Survival / Q000528:radiation effects; D004249:DNA Damage; D053843:DNA Mismatch Repair; D004256:DNA Polymerase I / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000648:ultrastructure; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D004789:Enzyme Activation; D016368:Frameshift Mutation; D016296:Mutagenesis; D009154:Mutation; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D054875:Ubiquitination / Q000235:genetics; D014466:Ultraviolet Rays
D004277:DNA, Single-Stranded; D018809:Proliferating Cell Nuclear Antigen; D029701:Saccharomyces cerevisiae Proteins; D004256:DNA Polymerase I; D004259:DNA-Directed DNA Polymerase