Structural study of immunoaffinity-purified DNA polymerase α-DNA primase complex from calf thymus

Tamai, Katsuyuki; Kojima, Kiyohide; Hanaichi, Takamasa; Masaki, Shigeo; Suzuki, Motoshi; Umekawa, Hayato; Yoshida, Shonen

doi:10.1016/0167-4781(88)90122-4

Cited by 135 publications

(116 citation statements)

References 33 publications

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“…We also analyzed the kinetics of compound 5, and obtained similar results (data not shown). In the kinetic analysis, poly(dA)/oligo(dT) [12][13][14][15][16][17][18] and dTTP were used as the DNA template-primer and nucleotide substrate, respectively. Double-reciprocal plots of the results show that inhibition of pol α activity by compound 6 was noncompetitive with the DNA template and the nucleotide substrate.…”

Section: Resultsmentioning

confidence: 99%

“…Activities of DNA polymerases were measured by the methods described previously. 24,25) The substrates of DNA polymerases used were poly(dA)/oligo(dT) [12][13][14][15][16][17][18] and dTTP as template-primer DNA and nucleotide substrate, respectively. One unit of each DNA polymerase activity was defined as the amount of enzyme that catalyzes the incorporation of 1 nmol of deoxyribonucleotide triphosphate (i.e., dTTP) into the synthetic template-primers (i.e., poly(dA)/oligo(dT) [12][13][14][15][16][17][18] , A/T=2/1) in 60 min at 37°C under the normal reaction conditions for each of the enzymes.…”

Section: Methodsmentioning

confidence: 99%

“…DNA polymerase (pol α) was purified from calf thymus by immunoaffinity column chromatography as described previously. 16) Recombinant rat pol β was purified from E. coli JMpβ5 as described by Date et al 17) Human pol γ catalytic subunit gene was cloned into pFastBac. Histidine-tagged enzyme was expressed using BAC-TO-BAC HT Baculovirus Expression System according to the supplier's manual (Life Technologies, MD) and purified using ProBoundresin (Invitrogen Japan, Tokyo).…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, information concerning the structural characteristics of these inhibitors may provide valuable insight for the design of new anti-cancer agents. [12][13][14][15][16][17][18] were purchased from Amersham Biosciences (Buckinghamshire, UK). Supercoiled pUC19 plasmid DNA was obtained from TaKaRa (Tokyo).…”

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confidence: 99%

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A novel DNA topoisomerase inhibitor: dehydroebriconic acid, one of the lanostane‐type triterpene acids from Poria cocos

et al. 2004

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Traditional Chinese medicinal plants are a treasure house for screening novel inhibitors of DNA polymerases and DNA topoisomerases from mammals; in the present study, nine lanostanetype triterpene acids were found in sclerotium of Poria cocos. Among the nine compounds, only dehydroebriconic acid could potently inhibit DNA topoisomerase II (topo II) activity (IC50=4.6 μM), while the compound moderately inhibited the activities of DNA polymerases α, β, γ, δ, ɛ, η, iota;, κ and λ only from mammals, to similar extents. Another compound, dehydrotrametenonic acid, also showed moderate inhibitory effects against topo II (IC50=37.5 μM) and weak effects against all the polymerases tested. Both compounds showed no inhibitory effect against topo I, higher plant (cauliflower) DNA polymerase I (α‐like polymerase) or II (βlike polymerase), calf thymus terminal deoxynucleotidyl transferase, human immunodeficiency virus type‐1 reverse transcriptase, prokaryotic DNA polymerases such as the Klenow fragment of E. coli DNA polymerase I, Taq DNA polymerase and T4 DNA polymerase, or DNA metabolic enzymes such as T7 RNA polymerase, T4 polynucleotide kinase and bovine deoxyribonu‐clease I. These findings suggest that dehydroebriconic acid and dehydrotrametenonic acid should be designated as topo II‐preferential inhibitors, although they also moderately inhibited all the mammalian DNA polymerases tested. Both dehydrotrametenonic acid and dehydroebriconic acid could prevent the growth of human gastric cancer cells, and their LD50 values were 63.6 and 38.4 μM, respectively. The cells were halted at the G1 phase in the cell cycle. The relation between the structure of triterpene acids and their inhibitory activities is discussed.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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A novel DNA topoisomerase inhibitor: dehydroebriconic acid, one of the lanostane‐type triterpene acids from Poria cocos

et al. 2004

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“…Pol α was purifi ed from calf thymus by immuno-affi nity column chromatography as described by Tamai et al 10 . Recombinant rat pol β was purifi ed from Escherichia coli JMpβ5 as described by Date et al 11 .…”

Section: Enzymesmentioning

confidence: 99%

.BETA.-Sitosteryl (6'-O-linoleoyl)-glucoside of Soybean (Glycine max L.) Crude Extract Inhibits Y-Family DNA Polymerases

et al. 2010

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Sulfated glycoglycerolipid from archaebacterium inhibits eukaryotic DNA polymerase α, β and retroviral reverse transcriptase and affects methyl methanesulfonate cytotoxicity

et al. 1998

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Structural study of immunoaffinity-purified DNA polymerase α-DNA primase complex from calf thymus

Cited by 135 publications

References 33 publications

A novel DNA topoisomerase inhibitor: dehydroebriconic acid, one of the lanostane‐type triterpene acids from Poria cocos

A novel DNA topoisomerase inhibitor: dehydroebriconic acid, one of the lanostane‐type triterpene acids from Poria cocos

.BETA.-Sitosteryl (6'-O-linoleoyl)-glucoside of Soybean (Glycine max L.) Crude Extract Inhibits Y-Family DNA Polymerases

Sulfated glycoglycerolipid from archaebacterium inhibits eukaryotic DNA polymerase α, β and retroviral reverse transcriptase and affects methyl methanesulfonate cytotoxicity

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