Rapid isolation of genes from an indexed genomic library ofC. cinereus in a novelpab1+ cosmid

Bottoli, Alan P. F.; Kertesz-Chaloupková, K.; Boulianne, Robert P.; Granado, José; Aebi, Markus; ̈es, Ursula Ku

doi:10.1016/s0167-7012(98)00109-2

Cited by 22 publications

(21 citation statements)

References 46 publications

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“…The pab1 1 wild-type gene of C. cinerea present in the cosmid backbone was used as a selection marker. Cosmid DNAs from 60 pools of each 96-well microtiter dish-arranged E. coli clones, and from subpools and individual clones of microtiter dish 40 were isolated (Bottoli et al 1999).…”

Section: Methodsmentioning

confidence: 99%

An Essential Gene for Fruiting Body Initiation in the Basidiomycete Coprinopsis cinerea Is Homologous to Bacterial Cyclopropane Fatty Acid Synthase Genes

et al. 2006

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The self-compatible Coprinopsis cinerea homokaryon AmutBmut produces fruiting bodies without prior mating to another strain. Early stages of fruiting body development include the dark-dependent formation of primary hyphal knots and their light-induced transition to the more compact secondary hyphal knots. The AmutBmut UV mutant 6-031 forms primary hyphal knots, but development arrests at the transition state by a recessive defect in the cfs1 gene, isolated from a cosmid library by mutant complementation. A normal primordia phenotype was achieved when cfs11 was embedded at both sides in at least 4.0 kb of native flanking DNA. Truncations of the flanking DNA lead to reduction in transformation frequencies and faults in primordia tissue formation, suggesting that the gene is also acting at later stages of development. The cfs1 gene encodes a protein highly similar to cyclopropane fatty acid synthases, a class of enzymes shown in prokaryotes and recently in a plant to convert membrane-bound unsaturated fatty acids into cyclopropane fatty acids. In C. cinerea 6-031, the mutant cfs1 allele carries a T-to-G transversion, leading to an amino acid substitution (Y441D) in a domain suggested to be involved in the catalytic function of the protein and/or membrane interaction.

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Section: Methodsmentioning

confidence: 99%

An Essential Gene for Fruiting Body Initiation in the Basidiomycete Coprinopsis cinerea Is Homologous to Bacterial Cyclopropane Fatty Acid Synthase Genes

et al. 2006

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“…The galectin region encompassing cgl1 and cgl2 was isolated from a cosmid library of homokaryon AmutBmut (Bottoli et al, 1999). A 7068 bp BamHI fragment of galectin cosmid 27A9 harbouring both cgl1 and cgl2 genomic DNA was subcloned into pBluescript KSk (Stratagene) and sequenced completely on both strands (Microsynth).…”

Section: Methodsmentioning

confidence: 99%

“…Southern blot experiments (data not shown) and specific PCR analysis (Bottoli et al, 1999), revealed that (top left), the mycelium did not contain any differentiated structures, whereas at days 6 and 7, hyphal knots were present (top right). At days 8 and 9 (bottom right), initials and small primordia were seen that matured up to day 11 (bottom left) to the stage where karyogamy can be induced.…”

Section: The Two Galectins Are Expressed From Adjacent Locimentioning

confidence: 99%

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Fruiting body development in Coprinus cinereus: regulated expression of two galectins secreted by a non-classical pathway The GenBank accession number for the sequence reported in this paper is AF130360

et al. 2000

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Fruiting body formation in the basidiomycete Coprinus cinereus is a developmental process that occurs as a response of the mycelium to external stimuli. First, localized, highly branched hyphal structures (knots) are formed as a reaction to nutritional depletion. Hyphal-knot formation is repressed by light ; however, light signals are essential for the development of the hyphal knot into an embryonic fruiting body (primordium) as well as karyogamy, meiosis and fruiting body maturation. The role of the different environmental signals in the initial phases of fruiting body development was analysed. It was observed that two fungal galectins, Cgl1 and Cgl2, are differentially regulated during fruiting body formation. cgl2 expression initiated in early stages of fruiting body development (hyphal knot formation) and was maintained until maturation of the fruiting body, whereas cgl1 was specifically expressed in primordia and mature fruiting bodies. Immunofluorescence and immunoelectron microscopy studies detected galectins within specific fruiting body tissues. They localized in the extracellular matrix and the cell wall but also in membrane-bound bodies in the cytoplasm. Heterologous expression of Cgl2 in Saccharomyces cerevisiae indicated that secretion of this protein occurred independently of the classical secretory pathway.

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“…To perform the silencing constructs of atr1 and chk1 genes, we constructed a recipient plasmid carrying the gpdII promoter, an efficient constitutive promoter from Agaricus bisporus previously used in C. cinerea (Burns et al 2005); as a fungal transcriptional terminator we used the 39-UTR region from the hygromycin-resistant gene from the pNEB-Hyg plasmid, which is used in U. maydis for transformation (Castillo-Lluva et al 2004); and as a selectable marker we used the pab1 gene from C. cinerea, which encodes PABA synthase, necessary for para-aminobenzoic acid production (Bottoli et al 1999). pBS (+)KS plasmid was used as backbone.…”

Section: Rnai Proceduresmentioning

confidence: 99%

A DNA Damage Checkpoint Pathway Coordinates the Division of Dikaryotic Cells in the Ink Cap MushroomCoprinopsis cinerea

et al. 2013

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The fungal fruiting body or mushroom is a multicellular structure essential for sexual reproduction. It is composed of dikaryotic cells that contain one haploid nucleus from each mating partner sharing the same cytoplasm without undergoing nuclear fusion. In the mushroom, the pileus bears the hymenium, a layer of cells that includes the specialized basidia in which nuclear fusion, meiosis, and sporulation occur. Coprinopsis cinerea is a well-known model fungus used to study developmental processes associated with the formation of the fruiting body. Here we describe that knocking down the expression of Atr1 and Chk1, two kinases shown to be involved in the response to DNA damage in a number of eukaryotic organisms, dramatically impairs the ability to develop fruiting bodies in C. cinerea, as well as other developmental decisions such as sclerotia formation. These developmental defects correlated with the impairment in silenced strains to sustain an appropriated dikaryotic cell cycle. Dikaryotic cells in which chk1 or atr1 genes were silenced displayed a higher level of asynchronous mitosis and as a consequence aberrant cells carrying an unbalanced dose of nuclei. Since fruiting body initiation is dependent on the balanced mating-type regulator doses present in the dikaryon, we believe that the observed developmental defects were a consequence of the impaired cell cycle in the dikaryon. Our results suggest a connection between the DNA damage response cascade, cell cycle regulation, and developmental processes in this fungus. IN fungal cells, mating-the process equivalent to fertilizationbrings together two haploid nuclei in the same cytoplasm. It is generally thought that this process is essentially followed by nuclear fusion, resulting in a diploid nucleus that either enters meiosis immediately (as occurs in the fission yeast Schizosaccharomyces pombe) or is maintained and proliferates in the diploid state (as happens in the budding yeast Saccharomyces cerevisiae). However, in a large number of fungi, mating does not result in diploid nuclei. Instead, they form dikaryons, cells that contain one haploid nucleus from each mating partner sharing the same cytoplasm for a period of time without undergoing nuclear fusion or meiosis. These dikaryons continue to propagate, and eventually nuclear fusion will take place, which will be followed by meiosis, closing the sexual cycle (Brown and Casselton 2001).Dikaryon cell division, also called conjugate division, represents a big challenge to the cell since it has to ensure that each daughter dikaryon inherits a balance of each parental genome. For that, a complex cell cycle is required that involves distinct mechanisms to maintain heterokaryosis after cell division. For example, for a large number of Basidiomycota (e.g., the mushroom Coprinopsis cinerea), conjugate division includes the formation of a structure known as the clamp connection or clamp cell, as well as the sorting of one of the nuclei to this structure (Casselton 1978;Brown and Casselton 2001). In the...

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Rapid isolation of genes from an indexed genomic library ofC. cinereus in a novelpab1+ cosmid

Cited by 22 publications

References 46 publications

An Essential Gene for Fruiting Body Initiation in the Basidiomycete Coprinopsis cinerea Is Homologous to Bacterial Cyclopropane Fatty Acid Synthase Genes

An Essential Gene for Fruiting Body Initiation in the Basidiomycete Coprinopsis cinerea Is Homologous to Bacterial Cyclopropane Fatty Acid Synthase Genes

Fruiting body development in Coprinus cinereus: regulated expression of two galectins secreted by a non-classical pathway The GenBank accession number for the sequence reported in this paper is AF130360

A DNA Damage Checkpoint Pathway Coordinates the Division of Dikaryotic Cells in the Ink Cap MushroomCoprinopsis cinerea

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