Phosphorylation of the C subunit (p66) of human DNA polymerase δ

Lemmens, Laure; Urbach, Serge; Prudent, Renaud; Cochet, Claude; Baldacci, Giuseppe; Hughes, Patrick

doi:10.1016/j.bbrc.2007.12.083

Cited by 13 publications

(12 citation statements)

References 11 publications

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“…Like the Pol32 subunit, p68 possesses a conventional PIP motif at the extreme carboxyl terminal end ( Bruning & Shamoo, 2004 ), and accordingly, deletion of last 20 aa of p68 that encompass PIP sequences failed to bind to PCNA. Another study also revealed a mechanism that could modulate the interaction of p68 with PCNA by a protein kinase–mediated phosphorylation of Ser458 in the PIP-box 456 QV S ITGFF 463 ( Lemmens et al, 2008 ). Similarly, a 22-aa oligopeptide containing the PIP sequence 57 LIQMRPFL 64 of p50 was shown to bind PCNA by far-Western analysis ( Lu et al, 2002 ; Wang et al, 2011 ); however, mutational analysis in this motif to provide functional evidence is yet to be carried out.…”

Section: Discussionmentioning

confidence: 99%

Human DNA polymerase delta is a pentameric holoenzyme with a dimeric p12 subunit

et al. 2019

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Human DNA polymerase delta (Polδ), a holoenzyme consisting of p125, p50, p68, and p12 subunits, plays an essential role in DNA replication, repair, and recombination. Herein, using multiple physicochemical and cellular approaches, we found that the p12 protein forms a dimer in solution. In vitro reconstitution and pull down of cellular Polδ by tagged p12 substantiate the pentameric nature of this critical holoenzyme. Furthermore, a consensus proliferating nuclear antigen (PCNA) interaction protein motif at the extreme carboxyl-terminal tail and a homodimerization domain at the amino terminus of the p12 subunit were identified. Mutational analyses of these motifs in p12 suggest that dimerization facilitates p12 binding to the interdomain connecting loop of PCNA. In addition, we observed that oligomerization of the smallest subunit of Polδ is evolutionarily conserved as Cdm1 of Schizosaccharomyces pombe also dimerizes. Thus, we suggest that human Polδ is a pentameric complex with a dimeric p12 subunit, and discuss implications of p12 dimerization in enzyme architecture and PCNA interaction during DNA replication.

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Section: Discussionmentioning

confidence: 99%

Human DNA polymerase delta is a pentameric holoenzyme with a dimeric p12 subunit

et al. 2019

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“…However, the subunit POLD3 (p66) seems to be necessary for accumulation of the POLD1 (p125) catalytic core at sites of local damage. 39 Interestingly, Lemmens et al 40 demonstrated that a highly conserved region between amino acids 384 and 399 of p66 is phosphorylated, and that this is associated with the colocalization of p66 with the p125 subunit. A plausible hypothesis therefore is that mutations in POLD3 impair the interaction with POLD1 and thus the proofreading function of POLD1.…”

Section: Spectrum Of Additional Polymerase Gene Variantsmentioning

confidence: 99%

Frequency and phenotypic spectrum of germline mutations in POLE and seven other polymerase genes in 266 patients with colorectal adenomas and carcinomas

Spier

Holzapfel

Altmüller

et al. 2015

Intl Journal of Cancer

130

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In a number of families with colorectal adenomatous polyposis or suspected Lynch syndrome/HNPCC, no germline alteration in the APC, MUTYH, or mismatch repair (MMR) genes are found. Missense mutations in the polymerase genes POLE and POLD1 have recently been identified as rare cause of multiple colorectal adenomas and carcinomas, a condition termed polymerase proofreading‐associated polyposis (PPAP). The aim of the present study was to evaluate the clinical relevance and phenotypic spectrum of polymerase germline mutations. Therefore, targeted sequencing of the polymerase genes POLD1, POLD2, POLD3, POLD4, POLE, POLE2, POLE3 and POLE4 was performed in 266 unrelated patients with polyposis or fulfilled Amsterdam criteria. The POLE mutation c.1270C>G;p.Leu424Val was detected in four unrelated patients. The mutation was present in 1.5% (4/266) of all patients, 4% (3/77) of all familial cases and 7% (2/30) of familial polyposis cases. The colorectal phenotype in 14 affected individuals ranged from typical adenomatous polyposis to a HNPCC phenotype, with high intrafamilial variability. Multiple colorectal carcinomas and duodenal adenomas were common, and one case of duodenal carcinoma was reported. Additionally, various extraintestinal lesions were evident. Nine further putative pathogenic variants were identified. The most promising was c.1306C>T;p.Pro436Ser in POLE. In conclusion, a PPAP was identified in a substantial number of polyposis and familial colorectal cancer patients. Screening for polymerase proofreading mutations should therefore be considered, particularly in unexplained familial cases. The present study broadens the phenotypic spectrum of PPAP to duodenal adenomas and carcinomas, and identified novel, potentially pathogenic variants in four polymerase genes.

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“…The immediate target may be p68; phosphoproteomic studies have shown that p68 is phosphorylated at a minimum of six sites in vivo [Olsen et al, 2006]. These sites are in sequence contexts that indicate that they are sites for CK2 phosphorylation, and it has been shown that p68 is phosphorylated at multiple sites in vitro by CK2 [Gao et al, 2008;Lemmens et al, 2008]. The p68 subunit has a PIP-box (PCNA-interacting protein) for interaction with PCNA at its remote C-terminus ] and a conserved site in yeast for interaction with Pol a .…”

Section: Regulation Of Pol D Activity By Protein Phosphorylationmentioning

confidence: 99%

Regulation of human DNA polymerase delta in the cellular responses to DNA damage

Lee

Zhang

Lin

et al. 2012

Environ and Mol Mutagen

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The p12 subunit of polymerase delta (Pol d) is degraded in response to DNA damage induced by UV, alkylating agents, oxidative, and replication stresses. This leads to the conversion of the Pol d4 holoenzyme to the heterotrimer, Pol d3. We review studies that establish that Pol d3 formation is an event that could have a major impact on cellular processes in genomic surveillance, DNA replication, and DNA repair. p12 degradation is dependent on the apical ataxia telangiectasia and Rad3 related (ATR) kinase and is mediated by the ubiquitin-proteasome system. Pol d3 exhibits properties of an ''antimutator'' polymerase, suggesting that it could contribute to an increased surveillance against mutagenesis, for example, when Pol d carries out bypass synthesis past small base lesions that engage in spurious base pairing. Chromatin immunoprecipitation analysis and examination of the spatiotemporal recruitment of Pol d to sites of DNA damage show that Pol d3 is the primary form of Pol d associated with cyclobutane pyrimidine dimer lesions and therefore should be considered as the operative form of Pol d engaged in DNA repair. We propose a model for the switching of Pol d with translesion polymerases, incorporating the salient features of the recently determined structure of monoubiquitinated proliferating cell nuclear antigen and emphasizing the role of Pol d3. Because of the critical role of Pol d activity in DNA replication and repair, the formation of Pol d3 in response to DNA damage opens the prospect that pleiotropic effects may ensue. This opens the horizons for future exploration of how this novel response to DNA damage contributes to genomic stability. Environ. Mol. Mutagen. 53:683-698, 2012. V V C 2012 Wiley Periodicals, Inc.

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Phosphorylation of the C subunit (p66) of human DNA polymerase δ

Cited by 13 publications

References 11 publications

Human DNA polymerase delta is a pentameric holoenzyme with a dimeric p12 subunit

Human DNA polymerase delta is a pentameric holoenzyme with a dimeric p12 subunit

Frequency and phenotypic spectrum of germline mutations in POLE and seven other polymerase genes in 266 patients with colorectal adenomas and carcinomas

Regulation of human DNA polymerase delta in the cellular responses to DNA damage

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