title: Different types of interaction between PCNA and PIP boxes contribute to distinct cellular functions of Y-family DNA polymerases.
abstract: Translesion DNA synthesis (TLS) by the Y-family DNA polymerases Polη, Polι and Polκ, mediated via interaction with proliferating cell nuclear antigen (PCNA), is a crucial pathway that protects human cells against DNA damage. We report that Polη has three PCNA-interacting protein (PIP) boxes (PIP1, 2, 3) that contribute differentially to two distinct functions, stimulation of DNA synthesis and promotion of PCNA ubiquitination. The latter function is strongly associated with formation of nuclear Polη foci, which co-localize with PCNA. We also show that Polκ has two functionally distinct PIP boxes, like Polη, whereas Polι has a single PIP box involved in stimulation of DNA synthesis. All three polymerases were additionally stimulated by mono-ubiquitinated PCNA in vitro. The three PIP boxes and a ubiquitin-binding zinc-finger of Polη exert redundant and additive effects in vivo via distinct molecular mechanisms. These findings provide an integrated picture of the orchestration of TLS polymerases.
authors.affiliation: Department of Genome Dynamics, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan Department of Toxicogenomics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Masuda
authors.affiliation: Department of Genome Dynamics, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Kanao
authors.affiliation: Department of Genome Dynamics, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kaji
authors.affiliation: Department of Gene Information, Institute for Virus Research, Kyoto University, Sakyo-ku, Kyoto 606-8517, Japan Department of Life Science, Graduate School of Science, Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo 171-8588, Japan.
authors.forename: Haruo
authors.initials: H
authors.lastname: Ohmori
authors.affiliation: Department of Life Science, Graduate School of Science, Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo 171-8588, Japan.
authors.forename: Fumio
authors.initials: F
authors.lastname: Hanaoka
authors.affiliation: Department of Genome Dynamics, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan masutani@riem.nagoya-u.ac.jp.
authors.forename: Chikahide
authors.initials: C
authors.lastname: Masutani
mesh_terms: D020816:Amino Acid Motifs; D002460:Cell Line; D004247:DNA / Q000096:biosynthesis*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009154:Mutation; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D054730:Protein Interaction Domains and Motifs; D054875:Ubiquitination; D000094769:DNA Polymerase iota; D000099270:Y-Family DNA Polymerases
chemical_list: D004247:DNA; D004259:DNA-Directed DNA Polymerase; D018809:Proliferating Cell Nuclear Antigen; D000094769:DNA Polymerase iota; C404302:POLK protein, human; D000099270:Y-Family DNA Polymerases; C112338:Rad30 protein; C000722070:POLI protein, human