Chromatin-Associated DNA Polymerase Activity in Meiotic Cells of Lily and Mouse;its Stimulation by Meiotic Helix-Destabilizing Protein

Sakaguchi, Kengo; Hotta, Yasuo; Stern, Herbert

doi:10.1247/csf.5.323

Cited by 83 publications

(59 citation statements)

References 24 publications

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“…Only recently, the ,B-like DNA polymerase has been reported in the slime mold Dictyostelium (1) and Physarum polycephalum (14) and in the protozoan Tetrahymena (4). In plants, the ,B-like enzyme has been found in Lilium (13) and cauliflower (3). The production of 13-like DNA polymerase is less than 10%o of the total enzyme in Dictyostelium somatic cells, it is limited to late log phase in Tetrahymena, and it is produced in excess of the a-like enzyme in meiotic cells in cauliflower and Lilium sporocytes.…”

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confidence: 99%

Meiosis in Coprinus: characterization and activities of two forms of DNA polymerase during meiotic stages.

Sakaguchi¹,

Lu²

1982

Mol. Cell. Biol.

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Two forms of DNA polymerase have been studied in the basidiomycete Coprinus. DNA polymerase from basidiocarp tissues at zygotene-pachytene stage has been purified 3,500-fold and defined as DNA polymerase b by virtue of its insensitivity to N-ethylmaleimide and by its low molecular weight (76,000). This enzyme has optimal activity at pH 7.0 to 7.5, at 200 mM KCI, and at 25C incubation temperature. It can use polycytidylic acid-oligo(dG)12_18 as template primer in addition to homodeoxypolymers. The DNA polymerase a is mainly produced in the exponentially growing mycelium. It is sensitive to N-ethylmaleimide and has a temperature optimum at 35°C. At the premeiotic S phase, activities from both polymerase a and polymerase b are found in cell-free extracts. The b enzyme is the only DNA polymerase produced during meiotic prophase. Its assayable activity exhibits two peaks, one at premeiotic S stage and one at pachytene. It is possible that DNA polymerase b is responsible for pachytene repairs involved in recombination.In recent years, we have been interested in the synchronous meiotic system of Coprinus (8,9a,11) and in particular, the DNA repair synthesis and its relationship to genetic recombination (10). In this connection, it is prudent to examine the activities of DNA polymerases during meiotic and premeiotic stages. Our finding has been most interesting, as this represents the first detailed characterization of meiotic DNA polymerase in the higher basidiomycetes.The presence of several distinct types of DNA-dependent DNA polymerases has been well documented; these include I, II, and III for bacteria (5) and alpha, beta, and gamma for mammalian cells (17). The same information is lacking in the lower eucaryotes, especially the higher fungi. As recently as 1977, the lower eucaryotes were believed not to have DNA polymerase , (2). Only recently, the ,B-like DNA polymerase has been reported in the slime mold Dictyostelium (1) and Physarum polycephalum (14) and in the protozoan Tetrahymena (4). In plants, the ,B-like enzyme has been found in Lilium (13) and cauliflower (3). The production of 13-like DNA polymerase is less than 10%o of the total enzyme in Dictyostelium somatic cells, it is limited to late log phase in Tetrahymena, and it is produced in excess of the a-like enzyme in meiotic cells in cauliflower and Lilium sporocytes. It is possible that the 1-like DNA polymerase has an important physiological function to play in meiotic processes and is therefore primarily produced during meiosis. For the present paper, we took advantage of the synchronous meiotic system of Coprinus cinereus to test this possibility, and we present evidence that the 1-like enzyme (to be called DNA polymerase b) is the only DNA polymerase produced during meiotic prophase in C. cinereus. MATERIALS AND METHODSTerminology. The DNA polymerases of procaryotes are named polymerase I, II, and III, whereas those of mammalian cells are classified as polymerase a, 1, and y. In the higher fungi, a new notation may be in order. We propose for the ...

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confidence: 99%

Meiosis in Coprinus: characterization and activities of two forms of DNA polymerase during meiotic stages.

Sakaguchi¹,

Lu²

1982

Mol. Cell. Biol.

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“…Human pols ‰ and Â were purified by the nuclear fractionation of human peripheral blood cancer cells (Molt-4) using the second subunit of pols ‰ and Â-conjugated affinity column chromatography, respectively (15). Pol · from a higher plant, cauliflower inflorescence, was purified according to the methods outlined by Sakaguchi et al (16). The Klenow fragment of pol I from E. coli and HIV-1 reverse transcriptase (recombinant) were purchased from Worthington Biochemical Corp. (Freehold, NJ, USA).…”

Section: Methodsmentioning

confidence: 99%

Inhibitory effects of docosyl p-coumarate on DNA topoisomerase activity and human cancer cell growth

Mizushina

Nishimura²,

Takenaka³

et al. 2010

Int J Oncol

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Abstract. We previously found six compounds of alkyl pcoumarates from a composite plant Artemisia annua L., and chemically synthesized these compounds (cis-isomer of C20, C22 and C24, and trans-isomer of C20, C22 and C24 of pcoumarates are compounds 1-6, respectively). This report describes the inhibitory activities of these alkyl p-coumarates against DNA polymerase (pol), DNA topoisomerase (topo), and human cancer cell growth. Among the compounds tested, compounds 1 and 4 weakly inhibited repair-related pol ß activity, but no compound influenced the activity of replicative pol ·. Compounds 4-6 and compounds 2 and 5 were potent inhibitors of human topos I and II, respectively. Compounds 2, 4, 5 and 6 also suppressed the growth of human colon carcinoma cell line, HCT116, with or without p53, suggesting that cell growth inhibition had the same tendency as the inhibition of topos rather than pols. Compound 5 (docosyl p-coumarate), which was the strongest inhibitor of topo II and cancer cell growth in the compounds tested, halted HCT116 p53 +/+ cells in G2/M phases, and induced apoptosis, although this compound did not affect the cell cycle of HCT116 p53-/-cells. These results suggest that the effect of p53-dependent cell cycle arrest may be effective for topo inhibition by compound 5. From these findings, the action mode of alkyl pcoumarates as an anti-cancer agent is discussed.

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“…Fish pols α and δ were purified from the testes of cherry salmon (Oncorhynchus masou) (22). Pol I (α-like) from a higher plant, cauliflower inflorescence, was purified according to the methods outlined by Sakaguchi et al (23). Recombinant rice (Oryza sativa L. cv.…”

Section: Methodsmentioning

confidence: 99%

The specific inhibitory effect of demethoxydehydroaltenusin, a derivative of dehydroaltenusin, on mammalian DNA polymerase α

Mizushina¹

1998

Int J Mol Med

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Abstract. In the screening of selective inhibitors of eukaryotic DNA polymerases (pols), dehydroaltenusin from the fungus Acremonium sp. was found to be an inhibitor of pol α. The present study succeeded in chemically synthesizing dehydroaltenusin, and the compound strongly inhibited calf pol α activity and weakly suppressed rat pol ß activity, with IC 50 values of 0.68 and 64 μM, respectively. We purified or synthesized various slightly modified derivatives of dehydroaltenusin, and using these, investigated the relationship between chemical structure and the inhibitory effects. These results suggest that the ketone group at the 5'-position in dehydroaltenusin is essential for pol inhibitory activity, and the group at the 5-position is important for the specificity of pol α inhibition. Demethoxydehydroaltenusin was found to be the most specific pol α inhibitor among the prepared derivatives, and the IC 50 values for pols α and ß were 0.24 and 89 μM, respectively. This compound did not influence the activities of other replicative pols such as pols δ and ε, and also demonstrated no effect on pol α activity from another vertebrate, fish and a plant species. Demethoxydehydroaltenusin also had no influence on the other pols and DNA metabolic enzymes tested. Therefore, demethoxydehydroaltenusin is of interest as a mammalian pol α-selective inhibitor as a 'chemical knockout agent' in vitro and in vivo. IntroductionThe human genome encodes 16 DNA polymerases (pols) that control cellular DNA synthesis (1). Eukaryotic cells reportedly contain three replicative types: pols α, δ and ε, mitochondrial pol γ, and at least twelve repair types: pols ß, δ, ε, ζ, η, θ, ι, κ, λ, μ, and σ and REV1 (2). Selective inhibitors of each pol are useful tools and molecular probes for distinguishing pols and for clarifying their biological and in vivo functions (3). For example, aphidicolin is a selective inhibitor of both pol α and eukaryotic DNA replicative pols δ and ε, indicating that these pols are essential for DNA replication (4), and this inhibitor has been very useful for studying the DNA replication system (5); however, aphidicolin is not capable of distinguishing pols α, δ and ε.Therefore, we established an assay method to detect pol inhibitors, and have screened for natural sources of inhibitors for more than 10 years. An inhibitor was isolated that selectively influences the activity of mammalian pol α, dehydroaltenusin, from a fungus (Acremonium sp.) collected from fields in the vicinity of Noda city in Chiba prefecture, Japan (6). A total chemical synthesis method was established for dehydroaltenusin and succeeded in completely synthesizing the compound (7-9).Subsequently, the slightly modified derivatives were purified or chemically synthesized to examine the structural relationship between dehydroaltenusin and pols. Dehydroaltenusin and its analogues represent a group of potentially useful agents to examine the precise role of each pol in vivo, and to develop a drug design strategy for cancer chemotherapy agents. The compou...

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Chromatin-Associated DNA Polymerase Activity in Meiotic Cells of Lily and Mouse;its Stimulation by Meiotic Helix-Destabilizing Protein

Cited by 83 publications

References 24 publications

Meiosis in Coprinus: characterization and activities of two forms of DNA polymerase during meiotic stages.

Meiosis in Coprinus: characterization and activities of two forms of DNA polymerase during meiotic stages.

Inhibitory effects of docosyl p-coumarate on DNA topoisomerase activity and human cancer cell growth

The specific inhibitory effect of demethoxydehydroaltenusin, a derivative of dehydroaltenusin, on mammalian DNA polymerase α

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