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

Sakaguchi, Kengo; Lu, Benjamin C.

doi:10.1128/mcb.2.7.752

Cited by 39 publications

(33 citation statements)

References 18 publications

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“…4,1984 (27). In the vegetative phase of this organism, nearly all of the polymerase activity is accounted for by polymerase A, whereas in meiotic S phase, polymerase A and B are nearly equal and at pachytene, the polymerase B accounts for over 95% of the activity.…”

Section: Discussionmentioning

confidence: 99%

DNA polymerases, deoxyribonucleases, and recombination during meiosis in Saccharomyces cerevisiae.

Resnick¹,

Sugino²,

Nitiss³

et al. 1984

Mol. Cell. Biol.

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We utilized strains of Saccharomyces cerevisiae that exhibit high efficiency of synchrony of meiosis to examine several aspects of meiosis including sporulation, recombination, DNA synthesis, DNA polymerase I and II, and Mg2+-dependent alkaline DNases. The kinetics of commitment to intragenic recombination and sporulation are similar. The synthesis of DNA, as measured directly with diphenylamine, appears to precede the commitment to recombination. Both DNA polymerase I and H activities and total DNA-synthesizing activity in crude extracts increase two-to threefold before the beginning of meiotic DNA synthesis. Increases of 10-to 20-fold over mitotic levels are found for Mg2+-dependent alkaline DNase activity in crude extracts before and during the commitment to meiotic intragenic recombination. Of particular interest is the comparable increase in a nuclease under the control of the RAD52 gene; this enzyme has been identified by the use of antibody raised against a similar enzyme from Neurospora crassa. Since the RAD52 gene is essential for meiotic recombination, the nuclease is implicated in the high levels of recombination observed during meiosis. The effects observed in this report are meiosis specific since they are not observed in an aa strain.

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Section: Discussionmentioning

confidence: 99%

DNA polymerases, deoxyribonucleases, and recombination during meiosis in Saccharomyces cerevisiae.

Resnick¹,

Sugino²,

Nitiss³

et al. 1984

Mol. Cell. Biol.

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“…It seems likely that the Coprinus DNA ligase plays an importmt role in the meiotic process in concert with the meiosis-specific DNA polymerase. The meiotic prophase in which the DNA ligase might work actively is a period for two special meiotic events: chromosome pairing and crossing-over [32,491. The identfication of a corresponding DNA ligase gene, including cDNA cloning and sequence analysis, could clarify the situation.…”

Section: Discussionmentioning

confidence: 99%

“…Dikaryotic cultures (5026+5132) were prepared on sterile horse dung in culture dishes (90x50 mm) and incubated in absolute darkness at 35°C for 5 days, at which time the mycelium completely covered the culture medium. In order to synchronize all fruiting bodies, the arrest-release protocol [32] was used. The arrested cultures were released at 9:OO by returning them to a 25°C incubator with a 16 h light/8 h dark cycle.…”

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confidence: 99%

Characterization of DNA Ligase from the Fungus Coprinus cinereus

Matsuda

Sakaguchi

Tsukada

et al. 1996

European Journal of Biochemistry

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) -EJB 95 2128/4 DNA ligase was highly purified from the fungus Coprinus cinereus at the meiotic recombination stage, pachytene. The pachytene DNA ligase showed three polypeptides with molecular masses of 88, 84 and 80 kDa, as estimated by the [-"PIAMP-labeling assay. These three polypeptides were susceptible to reaction with an mAb against a 16-amino-acid sequence in human DNA ligase I, which is conserved in C-terminal regions of mammalian, vaccinia virus and yeast DNA ligases. Since rapidly purified preparations from fresh pachytene cells exhibited a single polypeptide of DNA ligase with a molecular mass of 88 kDa, the smaller polypeptides seemed to be limited-degradation products of the 88-kDa polypeptide during the isolation and purification procedures. K, values for ATP and (dT),, hybridized with (dA),, were 1.5 pM and 90 nM, respectively. This enzyme was capable of joining (dT),,] . (rA),3 and (rA),2-ls .(dT), as well as (dT),, . (dA),, and able to ligate blunt-ended DNA in the presence of poly(ethylene glycol) 6000. DNA ligases were also partially purified from zygotene cells at the meiotic pairing stage and mitotic mycelium cells. In their molecular macs, immuno-reactivity, K, value and substrate specificity, they were indistinguishable from pachytene DNA ligase. These results suggest that the fungus C.cinereus at the pachytene stage contains DNA ligase with a molecular mass of 88 kDa as a main or a single species, which is quite similar to DNA ligases from the zygotene and mycelium cells in molecular and catalytic properties.Keywords: DNA ligase; Coprinus cinereus ; meiosis ; mycelium.DNA ligase catalyzes the formation of phosphodiester bonds between adjacent 5'-phosphoryl and 3'-hydroxyl termini at single-strand or double-strand breaks of duplex DNA. It is therefore an essential element of the coordinated multienzyme processes of DNA replication, DNA excision repair, DNA doublestrand break repair and DNA recombination. Genetic studies in Escherichia coli and yeasts have demonstrated that a single species of DNA ligase is essential for DNA replication, repair and recombination [l -41. In contrast, mammalian and Drosophilu melanogaster embryo cells contain at least two different species of the enzyme [5-131. DNA ligase I activity and the gene expression are higher in rapidly dividing cells than in nonproliferating cells, implying a role for DNA ligase I in DNA replication [14-181. In fact, complementation of the conditional lethal phenotype of Succharonzyces cerevisiue cdc9 DNA ligase mutant by human DNA ligase I cDNA demonstrates that this enzyme is the key enzyme for joining Okazaki fragments during laggingstrand DNA synthesis [19]. Direct evidence has been obtained that the human cell line 46BR, which has two missense mutations in different alleles of the DNA ligase I gene, is defective not only in DNA replication but also in DNA excision repair In contrast to the animal kingdom, much less is known about DNA ligase in fungi and plants, though earlier reports showed the presence of ATP-dependent DNA...

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“…We have investigated meiosis-related protein factors using meiotic cells in the basidiomycete C. cinereus (Sakaguchi & Lu, 1982;Matsuda et al, 1996;Sawado & Sakaguchi, 1997;Nara et al, 1999;Hamada et al, 2002;Namekawa et al, 2003). We reported preliminarily that C. cinereus meiotic tissues at prophase I contained DNA ligase activities (Matsuda et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

DNA ligase IV from a basidiomycete, Coprinus cinereus, and its expression during meiosis

et al. 2003

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DNA ligase IV is thought to be involved in DNA double-strand break repair and DNA nonhomologous end-joining pathways, but these mechanisms are still unclear. To investigate the roles of DNA ligase IV from a biologically functional viewpoint, the authors studied its relationship to meiosis in a basidiomycete, Coprinus cinereus, which shows a highly synchronous meiotic cell cycle. The C. cinereus cDNA homologue of DNA ligase IV (CcLIG4) was successfully cloned. The 3?2 kb clone including the ORF encoded a predicted product of 1025 amino acid residues with a molecular mass of 117 kDa. A specific inserted sequence composed of 95 amino acids rich in aspartic acid and glutamic acid could be detected between tandem BRCT domains. The inserted sequence had no sequence identity with other eukaryotic counterparts of DNA ligase IV or with another aspartic acid and glutamic acid rich sequence inserted in C. cinereus proliferating cell nuclear antigen (CcPCNA), although the length and the percentages of aspartic and glutamic acids were similar. In addition, the recombinant CcLIG4 protein not only showed ATP-dependent ligase activity, but also used (dT) 16 /poly(dA) and (dT) 16 /poly(rA) as substrates, and had double-strand ligation activity, like human DNA ligase IV. Northern hybridization analysis and in situ hybridization indicated that CcLIG4 was expressed not only at the pre-meiotic S phase but also at meiotic prophase I. Intense signals were observed in leptotene and zygotene. Based on these observations, the possible role(s) of C. cinereus DNA ligase IV during meiosis are discussed.

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

Cited by 39 publications

References 18 publications

DNA polymerases, deoxyribonucleases, and recombination during meiosis in Saccharomyces cerevisiae.

DNA polymerases, deoxyribonucleases, and recombination during meiosis in Saccharomyces cerevisiae.

Characterization of DNA Ligase from the Fungus Coprinus cinereus

DNA ligase IV from a basidiomycete, Coprinus cinereus, and its expression during meiosis

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