Expression of flap endonuclease-1 during meiosis in a basidiomycete, Coprinus cinereus

Yamaguchi, Taiki; Namekawa, Satoshi H.; Hamada, Fumika N.; Kasai, Nobuyuki; Nara, Takayuki; Watanabe, Kei; Iwabata, Kazuki; Ishizaki, Takashi; Ishii, S; Koshiyama, Akiyo; Inagaki, Sachiyo; Kimura, Seisuke; Sakaguchi, Kengo

doi:10.1016/j.fgb.2004.01.008

Cited by 7 publications

(10 citation statements)

References 31 publications

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“…Developmental studies in A. bisporus are still hindered due to the time and containment issues that exist when studying a genetically modified strain. C. cinerea provides a model system for the studies of gene expression throughout mushroom development (47,59), and heterologous expression of the A. bisporus SPR1 promoter fusion is a further demonstration of the inkcap host utility as a model species.…”

Section: Discussionmentioning

confidence: 99%

“…The ink-cap mushroom, Coprinopsis cinerea (formerly Coprinus cinereus), is a well-studied homobasidiomycete (12,43,47) that forms an excellent model system for studies of gene expression at several levels of differentiation, particularly mushroom development and meiotic processes (46,59). It has been used as an object for studies of development (32), mainly because of its relatively short life cycle, which can be completed in the laboratory within 2 weeks (44).…”

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confidence: 99%

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Characterization of Serine Proteinase Expression in Agaricus bisporus and Coprinopsis cinerea by Using Green Fluorescent Protein and the A. bisporus SPR1 Promoter

Heneghan

Porta

Zhang

et al. 2009

Appl Environ Microbiol

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The Agaricus bisporus serine proteinase 1 (SPR1) appears to be significant in both mycelial nutrition and senescence of the fruiting body. We report on the construction of an SPR promoter::green fluorescent protein (GFP) fusion cassette, pGreen_hph1_SPR_GFP, for the investigation of temporal and developmental expression of SPR1 in homobasidiomycetes and to determine how expression is linked to physiological and environmental stimuli. Monitoring of A. bisporus pGreen_hph1_SPR_GFP transformants on media rich in ammonia or containing different nitrogen sources demonstrated that SPR1 is produced in response to available nitrogen. In A. bisporus fruiting bodies, GFP activity was localized to the stipe of postharvest senescing sporophores. pGreen_hph1_SPR_GFP was also transformed into the model basidiomycete Coprinopsis cinerea. Endogenous C. cinerea proteinase activity was profiled during liquid culture and fruiting body development. Maximum activity was observed in the mature cap, while activity dropped during autolysis. Analysis of the C. cinerea genome revealed seven genes showing significant homology to the A. bisporus SPR1 and SPR2 genes. These genes contain the aspartic acid, histidine, and serine residues common to serine proteinases. Analysis of the promoter regions revealed at least one CreA and several AreA regulatory motifs in all sequences. Fruiting was induced in C. cinerea dikaryons, and fluorescence was determined in different developmental stages. GFP expression was observed throughout the life cycle, demonstrating that serine proteinase can be active in all stages of C. cinerea fruiting body development. Serine proteinase expression (GFP fluorescence) was most concentrated during development of young tissue, which may be indicative of high protein turnover during cell differentiation.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Characterization of Serine Proteinase Expression in Agaricus bisporus and Coprinopsis cinerea by Using Green Fluorescent Protein and the A. bisporus SPR1 Promoter

Heneghan

Porta

Zhang

et al. 2009

Appl Environ Microbiol

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“…We have conducted a number of studies of 3R (DNA replication, repair, recombination) enzymes in C. cinereus meiosis. We have found that transcripts of 3R-related proteins, such as DNA polymerase a, proliferating cell nuclear antigen (PCNA), DNA ligase I, Flap endonuclease 1, DNA ligase IV and Lim15/Dmc1, are abundant at meiotic prophase I, and we have discussed the roles of 3R enzymes during meiosis (Hamada et al, 2002;Namekawa et al, 2003a, b, c;Nara et al, 1999;Yamaguchi et al, 2004). These studies were made possible by the fact that large populations of synchronous meiotic cells can be obtained from C. cinereus to do biochemical analysis.…”

Section: Introductionmentioning

confidence: 99%

Knockdown of LIM15/DMC1 in the mushroom Coprinus cinereus by double-stranded RNA-mediated gene silencing

et al. 2005

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The basidiomycete Coprinus cinereus has many advantages as a model organism for studying sexual development and meiosis, but it has been difficult to investigate using reverse-genetics methods, such as gene disruption by homologous recombination. Here, gene repression by dsRNA-mediated gene silencing was tried as an alternative method for reverse-genetics studies. It was shown that transformation of the LIM15/DMC1 dsRNA expression construct (LIM15dsRNA) resulted in genomic insertion of LIM15dsRNA and paucity of the LIM15/DMC1 transcript. First, LIM15dsRNA was transformed into the homothallic strain AmutBmut to generate a homozygote in which both nuclei had a copy of LIM15dsRNA. The LIM15/DMC1-repressed strain showed abnormal homologous chromosome synapsis during meiosis. Basidiospore production was reduced to 16 % by the induction of dsRNA. However, approximately 60 % of basidiospores were viable. Next, a heterozygote was generated in which one nucleus had a copy of LIM15dsRNA. The phenotype was similar to that of the homozygote. These results are not only the first demonstration of dsRNA-mediated gene silencing in a member of the homobasidiomycete fungi, to which 90 % of mushroom species belong, but also the first successful use of a reverse-genetics approach in C. cinereus research.

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“…Not only is its meiotic cell cycle long and naturally synchronous, but large populations of synchronous meiotic cells are also easy to obtain (Li et al 1999;Kües 2000;Kamada 2002;Casselton and Olesnicky 1998). These advantages have enabled us to isolate and characterize many synapsis-related proteins in C. cinereus (Nara et al 1999(Nara et al , 2001Hamada et al 2007;Ishizaki et al 2002;Namekawa et al 2003a, b, c;Yamaguchi et al 2004;Iwabata et al 2005;Koshiyama et al 2006;Sakamoto et al 2007;Sakaguchi et al 2007) and to demonstrate their roles in vivo.…”

Section: Introductionmentioning

confidence: 99%

Coprinus cinereus Mer3 is required for synaptonemal complex formation during meiosis

et al. 2008

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Mer3 is an evolutionarily conserved DNA helicase that has crucial roles in meiotic recombination and crossover formation. We have identified the MER3 homolog in Coprinus cinereus (Ccmer3) and show that it is expressed in zygotene and pachytene meiocytes. Immunostaining analysis indicated that CcMer3 was localized on chromosomes at zygotene and pachytene and CcMer3 foci were more frequent on paired than unpaired chromosomes. We generated a C. cinereus mer3 mutant (#1) and found that it showed abnormal meiosis progression and underwent apoptosis after prophase I. Basidiospore production in #1 was reduced to 0.8% of the wild-type level; the spores showed slower germination at 25 degrees C but were similar to the wild type at 37 degrees C. Electron microscopic analysis of chromosome spreads revealed that axial elements were formed in the mutant but that synapsis was defective, resulting in a reduction in spore production. Our results demonstrate that CcMer3 is required for synaptonemal complex formation after axial elements align and is thus essential for homologous synapsis.

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Expression of flap endonuclease-1 during meiosis in a basidiomycete, Coprinus cinereus

Cited by 7 publications

References 31 publications

Characterization of Serine Proteinase Expression in Agaricus bisporus and Coprinopsis cinerea by Using Green Fluorescent Protein and the A. bisporus SPR1 Promoter

Characterization of Serine Proteinase Expression in Agaricus bisporus and Coprinopsis cinerea by Using Green Fluorescent Protein and the A. bisporus SPR1 Promoter

Knockdown of LIM15/DMC1 in the mushroom Coprinus cinereus by double-stranded RNA-mediated gene silencing

Coprinus cinereus Mer3 is required for synaptonemal complex formation during meiosis

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