DNA polymerases alpha, delta, and epsilon: three distinct enzymes from HeLa cells.

Syväoja, Juhani E.; Suomensaari, Susanna; Nishida, Craig; Goldsmith, Jeffrey S.; Chui, Gloria; Jain, Sanjay; Linn, Stuart

doi:10.1073/pnas.87.17.6664

Cited by 158 publications

(100 citation statements)

References 33 publications

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“…Earlier skepticism that Pol d was simply Pol a contaminated with a 3 0 -5 0 exonuclease activity was dispelled by the demonstration that both the polymerase and exonuclease activities were associated with the p125 catalytic subunit [Lee et al, 1980;Lee et al, 1984]. A second form of ''Pol d'' with a larger catalytic subunit was discovered and is now known as Pol e [Syvaoja et al, 1990;Pospiech and Syvaoja, 2003;Pursell and Kunkel, 2008]. Molecular cloning of p125 showed that the catalytic core was highly conserved in evolution from T4 bacteriophage to human Hao et al, 1992;Yang et al, 1992], and Pol d is now classified as a member of the B family polymerases.…”

Section: Introductionmentioning

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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Section: Introductionmentioning

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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“…Although the 3'?5' exonuclease activity of the wt p53 protein might provide a biochemical basis for the direct involvement of this molecule in the repair of certain types of DNA damage, the biological relevance of this enzyme activity remains elusive. The observations that p53 protein is co-localized with the DNA replication machinery (Gannon and Lane, 1987;Cox et al, 1995) and that mammalian DNA pol a, unlike pol d and pol e, lacks the 3'?5' exonuclease activity for proofreading (Syvaoja et al, 1990;Bambara and Jessee, 1991) provide hints that link p53 with DNA replication ®delity. Because the misincorporation of non-complementary nucleotides during DNA replication represents a major mechanism of gene mutation, prompt removal of the misincorporated nucleotides from DNA is critical for genomic stability.…”

Section: Introductionmentioning

confidence: 99%

Excision of mismatched nucleotides from DNA: a potential mechanism for enhancing DNA replication fidelity by the wild-type p53 protein

Huang

1998

Oncogene

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The tumor suppressor p53 plays a critical role in the regulation of the cell cycle and the maintenance of genetic stability. The 3'?5' exonuclease activity of p53 has recently been recognized as a novel biochemical function of this molecule, but the biological signi®cance of this activity remains elusive. Using an in vitro DNA replication assay with puri®ed human DNA polymerases, p53 protein, and de®ned DNA primer/templates, we demonstrated that the wild-type (wt) p53 protein, but not the mutant p53 protein, speci®cally enhanced the DNA replication ®delity of polymerase (pol) a, an enzyme that lacks 3'?5' exonuclease activity. The misincorporation of non-complementary deoxynucleotides into DNA by pol a was substantially decreased by p53. In contrast, wt p53 showed no signi®cant e ect on replication ®delity or the rates of DNA synthesis by human pol e or the bacterial enzyme pol I. Inhibition of 3'?5' exonuclease activity by guanosine monophosphate (GMP) abolished the ability of p53 to enhance the replication ®delity of pol a. Quantitative analyses revealed that the 3'?5' exonuclease activity of p53 e ectively removed mismatched nucleotides from DNA in preference over matched nucleotides. Furthermore, study in intact cells revealed that the wt p53 protein was co-localized with DNA synthesis activity in S phase cells. These results suggest a possibility that the 3'?5' exonuclease of the wt p53 protein might provide the proof-reading function for DNA pol a. The preferential excision of mismatched nucleotides from the replicating DNA strand appears to be a potential biochemical mechanism by which p53 enhances DNA replication ®delity and thereby helps to maintain genomic integrity.

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“…Similarly, it has been shown that SV40 DNA replication is also pol ⑀-independent (23), but this polymerase might be required for the chromosomal DNA replication in vivo (33). It has been shown that pol ⑀ is one of the replicative polymerases in HeLa cells (35). Studies from several laboratories suggest that pol ⑀ is involved in the cell cycle checkpoint control and may function to ensure accurate DNA synthesis (31, 34, 70, 71).…”

Section: Figmentioning

confidence: 99%

“…Unlike pol ␦, the role of pol ⑀ remains unclear. Pol ⑀ is a highly processive enzyme that has been implicated in the genomic DNA replication, DNA repair, recombination, as well as the S phase checkpoint control (31)(32)(33)(34)(35)(36).…”

mentioning

confidence: 99%

Modulation of DNA Synthesis in Saccharomyces cerevisiae Nuclear Extract by DNA Polymerases and the Origin Recognition Complex

Mitkova

Biswas-Fiss

Biswas

2005

Journal of Biological Chemistry

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We have analyzed the modulation of DNA synthesis on a supercoiled plasmid DNA template by DNA polymerases (pol), minichromosome maintenance protein complex (Mcm), topoisomerases, and the origin recognition complex (ORC) using an in vitro assay system. Antisera specific against the four-subunit pol ␣, the catalytic subunit of pol ␦, and the Mcm467 complex each inhibited DNA synthesis. However, DNA synthesis in this system appeared to be independent of pol⑀. Consequently, DNA synthesis in the in vitro system appeared to depend only on two polymerases, ␣ and ␦, as well as the Mcm467 DNA helicase. This system requires supercoiled plasmid DNA template and DNA synthesis absolutely required DNA topoisomerase I. In addition, we also report here a novel finding that purified recombinant six subunit ORC significantly stimulated the DNA synthesis on a supercoiled plasmid DNA template containing an autonomously replicating sequence, ARS1.Studies on the replication of genomes of bacteriophages, plasmids, viruses, and prokaryotic and eukaryotic cells have established general mechanisms of DNA replication (1). The eukaryotic origins of DNA replication were first discovered in the budding yeast, Saccharomyces cerevisiae, and were named autonomously replicating sequences (ARS) 1 (2-4). Many yeast ARS elements have been characterized in detail, which has allowed the discovery of many cis-acting replication proteins and regulators of DNA replication (5, 6). The discovery by Bell and Stillman (7,8) that the in vivo initiation of DNA replication in S. cerevisiae requires a six subunit origin recognition complex (ORC) is a major advancement in understanding the mechanism of eukaryotic DNA replication. In the G 1 phase of the cell cycle, Cdc6p, Cdt1p, minichromosome maintenance (Mcm), and Cdc45p proteins bind sequentially to DNA to form a prereplication complex (pre-RC) (9 -11). At the G 1 /S boundary, S phase-specific cyclin-dependent protein kinases (Cdks) and particularly the Cdc7/Dbf4 kinase transform the pre-RC into an active replication complex (10 -12). Finally, the initiation coincides with the association of the polymerase ␣/primase (pol ␣/primase) complex to the RPA-coated unwound origins (10). S and M phase-specific Cdks block the re-binding of Mcm to the chromatin and prevent a new round of initiations until mitosis (13).The Mcm protein family that is required for DNA replication in eukaryotes consists of six proteins (Mcm 2-7) with highly conserved amino acids between the six polypeptides. All of these proteins are essential for the cell viability (13, 14). The Mcm467 proteins form a relatively stable core complex and other Mcm proteins are loosely associated to it (14 -17). It has been demonstrated that a DNA helicase activity is associated with the Mcm467 complex, suggesting that this complex is involved in the initiation of DNA replication as a DNA unwinding enzyme (18,19). Furthermore, Mcm467 remains associated with the replication fork during the elongation step of DNA replication (20). Among a number of candidate D...

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DNA polymerases alpha, delta, and epsilon: three distinct enzymes from HeLa cells.

Cited by 158 publications

References 33 publications

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

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

Excision of mismatched nucleotides from DNA: a potential mechanism for enhancing DNA replication fidelity by the wild-type p53 protein

Modulation of DNA Synthesis in Saccharomyces cerevisiae Nuclear Extract by DNA Polymerases and the Origin Recognition Complex

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