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At least 55 records · Page 3Linked to original sources
PubMed1973

Changes in isozyme patterns between monokaryons and dikaryons of a bipolar Coprinus.

Proteins and isozymes of several different classes of enzymes in partially purified protein extracts of monokaryons, dikaryons, and monokaryon mixtures of a bipolar Coprinus sp. were separated on polyacrylamide gels by slab electrophoresis. Differences in protein and isozyme spectra were correlated with the operation of the incompatibility factors and with the results of Wang and Raper on Schizophyllum. It was concluded that the shift from monokaryon to dikaryon mediated a major change in the nature, quantity, or distribution of the proteins of this Coprinus sp.

Aminopeptidases↗
PubMed2002

The pab1 gene of Coprinus cinereus encodes a bifunctional protein for para-aminobenzoic acid (PABA) synthesis: implications for the evolution of fused PABA synthases.

The pab1 gene of the basidiomycete Coprinus cinereus encodes PABA synthase, necessary for para-aminobenzoic acid production. The C. cinereus protein is bifunctional with an N-terminal glutamine amidotransferase domain and a C-terminal chorismate amination domain. In most bacteria, these two functions are encoded in separate genes (e.g., pabA and pabB of E. coli). Fused PABA synthases have so far been detected in actinomycetes, Plasmodium falciparum, fungi and Arabidopsis thaliana. Phylogenetic analysis shows that the fused PAB sequences form a tight group that also includes uncharacterized PabB homologues from several bacteria. Unfused bacterial PabA proteins group with the glutamine amidotransferase subunits of bacterial anthranilate synthases, independent of organismal systematics, indicating a complex and perhaps independent evolutionary origin. In contrast, unfused PabB group and fused PabA/B proteins form a monophyletic group on a branch separate from the chorismate amination subunits of anthranilate synthases, probably reflecting a need for recognition of different positions in the common substrate chorismate.

4-Aminobenzoic Acid↗
PubMed2002

Molecular genetics of sexual development in the mushroom Coprinus cinereus.

Sexual development in the mushroom Coprinus cinereus is under the control of two mating type loci, A and B. When two haploid homokaryons with compatible alleles at both A and B loci are mated, the coordinated activities of A- and B-regulated pathways lead to formation of a mycelium termed the dikaryon, in which the two nuclei from the mating partners pair in each cell without fusing. The dikaryon is a prolonged mycelial stage that can be induced to develop a multicellular structure, the mushroom, under proper environmental conditions. The two nuclei fuse in specialized cells on the mushroom and immediately undergo meiosis to complete the sexual life cycle. It has been established recently that the A genes encode two classes of homeodomain proteins while the B genes encode pheromones and their receptors. More recently, molecular genetics has been used to reveal genes that work downstream of the mating type genes to regulate dikaryon formation, mushroom morphogenesis, and meiosis.

Coprinus↗
PubMed1994

A chimeric homeodomain protein causes self-compatibility and constitutive sexual development in the mushroom Coprinus cinereus.

The A mating type genes of the mushroom Coprinus cinereus encode two classes of putative transcription factor with distinctive homeodomain motifs (HD1 and HD2). A successful mating brings together different allelic forms of these genes and this triggers part of a developmental sequence required for sexual reproduction. In this report we provide evidence that this developmental programme is promoted by a physical interaction between the two classes of homeodomain protein. Rare dominant mutations conferring self-compatibility map to the A locus and result in constitutive operation of the A-regulated developmental pathway. Our molecular analysis of one of these mutations shows that it has generated a chimeric gene by inframe fusion of an HD2 and an HD1 gene. Fusion has overcome the normal incompatibility between two proteins coded by genes of the same A locus and generated a protein that is sufficient to promote development in the absence of any other active A mating type genes. The fusion protein retains most of the HD2 sequence, but only the C-terminal part of the HD1 protein. It has only the HD2 homeodomain motif as a potential DNA binding domain fused to an essential C-terminal region of the HD1 protein, which in a normal HD1-HD2 protein complex may be the major activation domain.

Amino Acid Sequence↗
PubMed1994

Enzymatic removal of selected aromatic contaminants from wastewater by a fungal peroxidase from Coprinus macrorhizus in batch reactors.

The use of enzymes such as horseradish peroxidase (HRP) for degrading or removing toxic organics from synthetic wastewater has been demonstrated previously. Potential alternatives to HRP are other peroxidases, various ligninases, haloperoxidases and laccases. Results of this study indicate that a fungal peroxidase from Coprinus macrorhizus (CMP) has the capability to catalyze the same reactions as HRP. Similarly, in batch reactors the trend and removal efficiency of aromatic compounds by CMP from synthetic wastewater depend on the nature of the compound.

Biotechnology↗
PubMed2000

Characterization of interaction of C- and N-terminal domains in LIM15/DMC1 and RAD51 from a basidiomycetes, Coprinus cinereus.

Both LIM15/DMC1 and RAD51 are thought to be essential for meiosis in which homologous chromosomes pair and recombine. The primary purpose of the present study was to investigate the homotypic and heterotypic interactions among their terminal domains. We prepared cDNAs and recombinant proteins of the full-length, N-terminal, and the C-terminal domains of LIM15/DMC1 (CoLIM15) and RAD51 (CoRAD51) from the basidiomycete Coprinus cinereus. In both two-hybrid assay in vivo and pull-down assay in vitro, either CoLim15 or CoRad51 interacted homotypically between the C-terminal domains, respectively, but no heterotypic interaction was observed between CoLim15 and CoRad51. The N-terminal domain of CoLim15 bound to ssDNA and dsDNA, while the C-terminal domain of CoRad51 appeared to interact weakly with ssDNA. Based on these results, the interaction among the strand-exchange proteins and meiosis was discussed.

Cell Cycle Proteins↗
PubMed1997

Expression of two closely linked hydrophobin genes of Coprinus cinereus is monokaryon-specific and down-regulated by the oid-1 mutation.

A protein with characteristic properties of a fungal hydrophobin (CoH1) was isolated from the monokaryotic stage of the basidiomycete Coprinus cinereus. A cosmid clone containing the corresponding gene (coH1) was identified using a cDNA probe derived by RT-PCR. Hybridization and sequence analysis identified a second gene, coH2, just 4.1 kb downstream of coH1 encoding a hydrophobin (CoH2) with 64% sequence identity. Both coH1 and coH2 are subject to developmental regulation. They are expressed in vegetative monokaryotic cells but not in the asexual oidia produced on the surface of monokaryons. Transcripts of the genes were barely detected in dikaryotic mycelium and were absent from fruit bodies. Loss of aerial growth due to a mutation known as oid-1 was correlated with lack of both hydrophobins.

Amino Acid Sequence↗
PubMed2000

A mutation in the eln2 gene encoding a cytochrome P450 of Coprinus cinereus affects mushroom morphogenesis.

A dominant mutation of the elongationless2 (eln2) gene of the mushroom Coprinus cinereus (eln2-1) affects pattern formation in the development of fruit body primordia, causing dumpy primordia which culminate in mature fruit bodies with short stipes. Phenotypic analyses revealed disturbance of tissue organization and cell morphogenesis in the primordial shaft and suggested that the defects in the primordial shaft interfere with differentiation of the stipe tissue in the mutant primordia. The eln2 locus was assigned to chromosome XII and the eln2-1 mutant gene was cloned from a chromosome XII-specific cosmid library as a genomic fragment that brings about the dominant mutant phenotype. The eln2 gene encodes a novel type of microsomal cytochrome P450 enzyme (CYP502) and is constitutively expressed. The eln2-1 mutation is a 4-bp deletion in the 3'-terminal region of the gene, leading to truncation of C-terminal 18 amino acids. We suggest that the truncation affects the activity of the CYP502 enzyme.

Amino Acid Sequence↗
PubMed2000

The control of meiosis progression in the fungus Coprinus cinereus by light/dark cycles.

Meiosis progression in Coprinus cinereus is controlled by light/dark cycles. Light is essential to propel basidia into karyogamy and light intensity determines the timing of meiotic events. The higher the light intensities, the faster the fruiting bodies enter karyogamy. The critical period when light has this influence is between 16 and 6 h before karyogamy. The control is highly stage specific. A 3-h dark period is essential for a Java dikaryon and the Japanese A(mut)B(mut) homokaryon to enter meiotic metaphase; without it the fruit body is permanently arrested at diffused diplotene. This arrest is light intensity-dependent (>20 hlx) and temperature-dependent (e.g., 27 degrees C). The placement of the dark period is very stage specific; it has no effect when placed before karyogamy stage. A dikaryon of London origin is light blind and able to complete meiosis under continuous high light regime. Fruiting bodies arrested under a continuous high light can be rescued by a 3-h dark treatment, but there is always an 8-h lag time to enter meiotic metaphase. It is possible that the dark effect signals cellular processes leading to division events. Cytological studies of arrested fruiting bodies showed that chromosomes are normal in meiotic prophase through pachytene and diplotene, but are unable to undergo chromosome condensation. Genetic crosses between a monokaryon of Java stock J6;5.4 and a monokaryon BL55 or H5 of London stock showed that light-blindness is dominant, and is controlled by a single Mendelian gene.

Circadian Rhythm↗
PubMed1993

Crystallization and X-ray diffraction analysis of recombinant Coprinus cinereus peroxidase.

Crystals suitable for an X-ray diffraction investigation have been obtained of recombinant Coprinus cinereus peroxidase expressed in Aspergillus oryzae. The crystals were grown by the hanging drop method with polyethylene glycol 6000 as the precipitant. A pH range from 6.2 to 8.0 and CaCl2 or MgCl2 present at a concentration of 0.35 M were essential for the crystal growth. A metastable monoclinic modification can be obtained under certain conditions, and with variations in temperature they are transformed into a stable orthorhombic modification. With CaCl2 as the additive, the unit cell dimensions were a = 74.9 A, b = 76.8 A and c = 128.2 A. With two peroxidase molecules per asymmetric unit, the solvent content is 49% (v/v). In the diffraction pattern, the reflections Okl are systematically very weak for k = 2n + 1. Combined with an analysis of the Patterson function, this showed that the two independent molecules are related by the pseudotranslational symmetry 0.29a + 0.5b. The possible space groups are P2(1)2(1)2(1) or P2(1)22(1) because of this pseudosymmetry. The crystals diffract to a resolution of 2.9 A.

Coprinus↗
PubMed1991

Targeted transformation in Coprinus cinereus.

We examined the influence of DNA form and size on the arrangement and genomic location of transforming DNA sequences in the basidiomycete Coprinus cinereus. Protoplasts with either single or double mutations in the tryptophan synthetase (TRP1) gene were transformed with cloned copies of this gene which contained only a single DNA strand, contained a specific single nick within the C. cinereus sequences (4.8 kb), contained a specific double-strand break, or contained an additional 35 kb of flanking genomic sequences. Gene replacement events were recovered when each DNA type was used. However, none of these substrates offers a substantial improvement in transformation or targeting frequency when compared to supercoiled circular DNA, which has allowed recovery of both gene replacements as well as homologous insertions in 5% of the transformants analyzed. The frequency of transformants carrying tandem insertions with multiple copies of the transforming DNA was reduced when single-stranded DNA was used, and increased when DNA containing double-strand breaks was used. These results have important implications for the efficient design of targeted transformation and co-transformation experiments.

Blotting, Southern↗
PubMed1993

The A mating-type genes of the mushroom Coprinus cinereus are not differentially transcribed in monokaryons and dikaryons.

The A mating type factor of Coprinus cinereus regulates part of a developmental sequence that leads to the conversion of the asexual monokaryon into the fertile dikaryon. The A42 factor is a complex of seven genes, at least four of which are involved in determining the specificity of mating interactions. In this report we show that the A42 genes are constitutively expressed in both monokaryons and dikaryons. This has important implications with respect to intracellular recognition of a compatible mating, which requires an interaction between proteins already present within the cells of the mating partners, and for the subsequent maintenance of dikaryotic growth.

Blotting, Northern↗
PubMed1994

A mating-type factors of Coprinus cinereus have variable numbers of specificity genes encoding two classes of homeodomain proteins.

We have identified the seven genes that constitute the A43 mating-type factor of Coprinus cinereus and compare the organisation of A43 with the previously characterised A42 factor. In both, the genes that trigger clamp cell development, the so-called specificity genes, are separated into alpha and beta loci by 7 kb of noncoding sequence and are flanked by homologous genes alpha-fg and beta-fg. The specificity genes are known to encode two classes of dissimilar homeodomain (HD1 and HD2) proteins and have different allelic forms which show little or no cross-hybridisation. By partial sequencing we identified a divergently transcribed HD1 (a1-2) and HD2 (a2-2) gene in the A43 alpha locus. a2-2 failed to elicit clamp cell development in three different hosts, suggesting that it is non-functional. a1-2 elicited clamp cells in an A42 host that has only an HD2 gene (a2-1) in its alpha locus, thus demonstrating that the compatible A alpha mating interaction is between an HD1 and an HD2 protein. The A43 beta locus contains three specificity genes, the divergently transcribed HD1 and HD2 genes b1-2 and b2-2 and a third HD1 gene (d1-1) that was shown by hybridisation and transformation analyses to be functionally equivalent to d1-1 in A42. An untranscribed footprint of a third A42 HD1 gene, c1-1, was detected between the A43 b2-2 and d1-1 genes by Southern hybridisation.

Amino Acid Sequence↗
PubMed1992

The acu-1 gene of Coprinus cinereus is a regulatory gene required for induction of acetate utilisation enzymes.

We have isolated a gene from Coprinus cinereus which cross-hybridises to the facA and acu-5 genes of Aspergillus nidulans and Neurospora crassa, respectively. These genes encode acetyl-CoA synthetase, an enzyme which is inducible by acetate and required for growth on acetate as sole carbon source. We have designated the C. cinereus gene acs-1 and have used transformation to demonstrate its functional homology to the ascomycete genes by complementation of an N. crassa acu-5 mutation. The acs-1 gene has never been identified by mutation; mutations leading to loss of acetyl-CoA synthetase function map to another gene, acu-1. Using Northern analyses we have shown that acu-1 has a regulatory function that is required for acetate-induced transcription of acs-1 and of another acetate utilisation gene, acu-7, the isocitrate lyase structural gene.

Acetates↗
PubMed1995

Chromosome dynamics in rad12 mutants of Coprinus cinereus.

We have characterized the phenotypes of three rad12 mutants of the basidiomycete Coprinus cinereus, which were isolated on the basis of sensitivity to ionizing radiation. Electron microscopic studies of meiotic nuclear spreads showed that all three rad12 mutants are defective in chromosomal synapsis. For rad12-1 and rad12-4, very limited assembly of the synaptonemal complex occurs. The phenotype of rad12-15 is less severe and longer stretches of synapsed chromosomes are formed. However, for all three alleles mutant nuclei arrest in a diffuse state with little synaptonemal complex structure. Observations made of spreads of acridine orange-stained meiotic nuclei correlated with the electron microscopic data. In rad12 strains, chromosomes condense but do not pair, and they later arrest in a decondensed state; very few rad12 cells enter metaphase I. Homozygous dikaryons of rad12 mutants produce fruiting bodies with significantly fewer basidiospores than are found in wild-type dikaryons. The viability of these spores is greatly reduced: all spores produced by rad12-1 and rad12-4 mushrooms fail to germinate, while only 16% of rad12-15 spores are viable. Recombination within the tract of the ribosomal RNA gene repeats was not significantly different in the mutants when compared with a wild-type congenic control. Quantitative measurements of oidial survival indicate that all three rad12 alleles are sensitive to gamma radiation but insensitive to UV radiation relative to wild-type strains.

Cell Nucleus↗