The mode of action of aphidicolin on DNA synthesis in isolated nuclei

Oguro, Mieko; Shioda, Masaki; Nagano, Hiroshi; Mano, Yoshitake; Hanaoka, Fumio; Yamada, Mami

doi:10.1016/0006-291x(80)91512-0

Cited by 51 publications

(20 citation statements)

References 13 publications

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“…6). The mode of inhibition of both polymerases by aphidicolin is competitive with deoxycytidine triphosphate (data not shown), as previously demonstrated for DNA polymerase a from sea urchin embryos (19), mouse myeloma (19), HeLa (18), and KB cells (12), and cultured rice cells hand, DNA polymerase y was completely resistant towards aphidicolin, even at 50 ug/ml, the highest concentration tested (Fig. 6).…”

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confidence: 83%

“…DISCUSSION DNA polymerase a plays a major role in the amplification of ribosomal RNA genes during oogenesis in X. laevis. This conclusion is derived from the observation that ribosomal DNA replication in isolated ovaries is completely inhibited by aphidicolin, a specific inhibitor of eucaryotic a-type DNA polymerases (12,16,18,19,24). In addition, we could show that the two aphidicolin-sensitive DNA polymerases isolated from ovaries of adult frogs are a-type polymerases, based on their preference for activated DNA as template, their inability to read (rA)n. (dT)1218, their inhibition by N-ethylmaleimide, and their relative high resistance towards dideoxy-TTP.…”

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confidence: 88%

“…To demonstrate that Xenopus DNA polymerase a is inhibited by aphidicolin as shown in a number of higher animal species (16,(18)(19)(20), DNA polymerases a and y were isolated from ovaries of adult frogs. The ovarian extract, obtained as described in Materials and Methods, was first bound to a DEAE-cellulose column and subsequently eluted with a potassium phosphate gradient.…”

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confidence: 99%

“…(dT)12 18 as a template. No evidence was found for the presence of DNA polymerase in the ovarian extract.…”

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confidence: 99%

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Participation of deoxyribonucleic acid polymerase alpha in amplification of ribosomal deoxyribonucleic acid in Xenopus laevis.

Zimmermann¹,

Weissbach²

1981

Mol. Cell. Biol.

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Aphidicolin, a known inhibitor of eucaryotic deoxyribonucleic acid (DNA) polymerase a, efficiently inhibited amplification of ribosomal DNA during oogenesis in Xenopus laevis. DNA polymerase a, but not DNA polymerase y, as isolated from ovaries, was sensitive to aphidicolin. DNA polymerase ,B was not detectable in Xenopus ovary extracts. Therefore, DNA polymerase a plays a major role in ribosomal ribonucleic acid gene amplification.During oogenesis in many different species, a large number of ribosomes are accumulated. These are needed for the extensive protein synthesis in the rapidly proliferating cells of the embryo. In some organisms, this process is facilitated by selective amplification of the genes that code for ribosomal ribonucleic acid (RNA) (9), thus providing additional templates for the massive ribosomal RNA synthesis which occurs during the development of the oocyte.In the South African clawed frog, Xenopus laevis, diploid somatic cells contain some 900 copies of the ribosomal RNA gene (2, 5) clustered together in the chromosomal nucleolus organizer regions (22). This repetitive deoxyribonucleic acid (DNA) contains, per repeat unit, the sequences for one copy each of 28S, 18S, and 5.8S ribosomal RNA and transcribed and nontranscribed spacers (2,5,7,17,26). During the early meiotic prophase in the oocyte (1, 6), the ribosomal RNA genes are amplified more than 1,000-fold and stored extrachromosomally in the nucleoplasn, forming hundreds of nucleoli (for review see reference 9).The mechanism of ribosomal DNA amplification in Xenopus proceeds by the selective replication of one of the repeat units of the ribosomal RNA genes, which exist in Xenopus as a multigene family whose members differ in the length of the nontranscribed DNA region (30,31). During amplification, a circularized copy of the ribosomal DNA is multiplied by a rollingcircle mechanism (13,23,31), producing multiple identical copies, tandemly arranged in circular and linear molecules (31). To date, it is not known which of the DNA polymerases is involved in ribosomal DNA amplification. We, therefore, investigated the role of DNA polymerase a in this process by using aphidicolin as a specific a polymerase inhibitor (14

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confidence: 83%

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confidence: 88%

Section: Downloaded Frommentioning

confidence: 99%

“…(dT)12 18 as a template. No evidence was found for the presence of DNA polymerase in the ovarian extract.…”

Section: Downloaded Frommentioning

confidence: 99%

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Participation of deoxyribonucleic acid polymerase alpha in amplification of ribosomal deoxyribonucleic acid in Xenopus laevis.

Zimmermann¹,

Weissbach²

1981

Mol. Cell. Biol.

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“…One must keep in mind not only that chemically synthesized nucleotide analogs can act as inhibitors of RT but also that there are several examples of highly specific enzyme inhibitors derived from natural sources, so there may be highly specific and effective natural inhibitors of RT that await detection. Aphidicolin, a specific inhibitor of DNA polymerase alpha (21), is an example of an inhibitor provided by naturally occurring organisms, e.g., fungi. Such substances are not necessarily thymidine derivatives and would obviously not be detected in a screening system with poly(rA)-oligo(dT) as the template primer.…”

Section: Discussionmentioning

confidence: 99%

Azidothymidine triphosphate is an inhibitor of both human immunodeficiency virus type 1 reverse transcriptase and DNA polymerase gamma

König

Behr

Löwer

et al. 1989

Antimicrob Agents Chemother

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The reverse transcriptase from human immunodeficiency virus type 1 was purified from the virus to near homogeneity. The enzyme was shown to possess both RNA-dependent and DNA-dependent DNA-synthesizing activity. Activated DNA as a heteropolymeric substrate was used as efficiently as was the homopolymeric substrate poly(rA)-oligo(dT). The Michaelis-Menten constants were determined for each of the four nucleotides needed to elongate a natural template primer. Azidothymidine triphosphate, a well-known inhibitor of the enzyme, inhibited the enzyme competitively with respect to dTTP and noncompetitively with respect to the other nucleotides. Azidothymidine triphosphate acted as an efficient inhibitor of cellular DNA polymerase gamma, whereas other enzymes of eucaryotic DNA metabolism, namely, DNA polymerase alpha-primase and DNA polymerase beta, were not inhibited. This finding may explain why some acquired immunodeficiency syndrome patients suffer side effects during azidothymidine therapy.

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Interaction of Epstein-Barr virus DNA polymerase with aphidicolin, phosphonoformate and 5?-GMP

Cheng

1988

Virus Genes

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Epstein-Barr virus (EBV)-specified DNA polymerase was purified from P3HR-1 cells, a Burkitt lymphoma EBV producer cell line, treated with phorbol-12,13-dibutyrate (PDB) and n-butyrate. Its inhibition by aphidicolin, phosphonoformate (PFA) and 5'-GMP was examined. Aphidicolin could inhibit EBV DNA polymerase competitively with respect to dATP and dCTP and noncompetitively with respect to dGTP and dTTP; whereas 5'-GMP was a noncompetitive inhibitor with respect to all four dNTPs. Combinations of aphidicolin and PFA, or PFA and 5'-GMP, produced a mutually exclusive inhibition pattern of EBV DNA polymerase that suggested that the binding sites of these compounds on the enzyme molecule are kinetically overlapping.

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The mode of action of aphidicolin on DNA synthesis in isolated nuclei

Cited by 51 publications

References 13 publications

Participation of deoxyribonucleic acid polymerase alpha in amplification of ribosomal deoxyribonucleic acid in Xenopus laevis.

Participation of deoxyribonucleic acid polymerase alpha in amplification of ribosomal deoxyribonucleic acid in Xenopus laevis.

Azidothymidine triphosphate is an inhibitor of both human immunodeficiency virus type 1 reverse transcriptase and DNA polymerase gamma

Interaction of Epstein-Barr virus DNA polymerase with aphidicolin, phosphonoformate and 5?-GMP

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