Interaction of the Aα Y and Z Mating-Type Homeodomain Proteins of Schizophyllum commune Detected by the Two-Hybrid System

Magae, Yumi; Novotny, Charles P.; Ullrich, Robert C.

doi:10.1006/bbrc.1995.1920

Cited by 36 publications

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“…When protein-protein interactions of homeodomain proteins in Schizophyllum commune were tested by using filter and liquid assays for ␤-galactosidase, moderate ␤-galactosidase activities (ϳ0.44 U/h per cell in yeast strain HF7c and Ͼ1.3 U/h per cell in yeast strain FY526) between two homeodomain proteins from different A␣ backgrounds were detected, confirming the ability of two different mating proteins to form a heterodimer (23). In our present research with yeast two-hybrid systems, we used an ␣-galactosidase assay, which is more sensitive than a ␤-galactosidase assay, to detect the Hox protein interaction.…”

Section: Vol 9 2010mentioning

confidence: 91%

“…On the basis of the homeodomain sequence, the mating-type proteins of the A locus are divided into two subgroups: HD1 and HD2 (20,21). When an HD1 protein from one mate heterodimerizes with an HD2 protein from the other mate to form a functional regulatory protein, sexual compatibility is intracellularly recognized, and the A developmental pathway is initiated (3,17,23).Few studies have examined the composition and function of mating-type loci in bipolar basidiomycetes. In a landmark study, Bakkeren and Kronstad (2) discovered that in bipolar fungus, Ustilago hordei, the A-and B-mating-type loci were fused into one nonrecombining mating-type region with two alleles.…”

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A -Mating-Type Gene Expression Can Drive Clamp Formation in the Bipolar Mushroom Pholiota microspora (Pholiota nameko)

Mukaiyama

Tachikawa

et al. 2010

Eukaryot Cell

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In the bipolar basidiomycete Pholiota microspora, a pair of homeodomain protein genes located at the A-mating-type locus regulates mating compatibility. In the present study, we used a DNA-mediated transformation system in P. microspora to investigate the homeodomain proteins that control the clamp formation. When a single homeodomain protein gene (A3-hox1 or A3-hox2) from the A3 monokaryon strain was transformed into the A4 monokaryon strain, the transformants produced many pseudoclamps but very few clamps. When two homeodomain protein genes (A3-hox1 and A3-hox2) were transformed either separately or together into the A4 monokaryon, the ratio of clamps to the clamplike cells in the transformants was significantly increased to ca. 50%. We therefore concluded that the gene dosage of homeodomain protein genes is important for clamp formation. When the sip promoter was connected to the coding region of A3-hox1 and A3-hox2 and the fused fragments were introduced into NGW19-6 (A4), the transformants achieved more than 85% clamp formation and exhibited two nuclei per cell, similar to the dikaryon (NGW12-163 ؋ NGW19-6). The results of real-time reverse transcription-PCR confirmed that sip promoter activity is greater than that of the native promoter of homeodomain protein genes in P. microspora. Thus, we concluded that nearly 100% clamp formation requires high expression levels of homeodomain protein genes and that altered expression of the A-mating-type genes alone is sufficient to drive true clamp formation.In basidiomycetous mushrooms, mating compatibility is controlled by one or two sets of multiple allelomorphic genes known as bipolar or tetrapolar mating systems, respectively (37). In tetrapolar mushrooms, such as Coprinopsis cinerea (5, 13), Laccaria bicolor (8,11,19), and Schizophyllum commune (10), the mating-type loci A and B, which are located on different chromosomes, regulate mating and clamp formation (9,15,27,28). The A locus comprises multigenes encoding homeodomain proteins, and the B locus comprises multigenes encoding pheromones and pheromone receptor proteins (6,13,21,25,26,30,31,32,35,36). On the basis of the homeodomain sequence, the mating-type proteins of the A locus are divided into two subgroups: HD1 and HD2 (20,21). When an HD1 protein from one mate heterodimerizes with an HD2 protein from the other mate to form a functional regulatory protein, sexual compatibility is intracellularly recognized, and the A developmental pathway is initiated (3,17,23).Few studies have examined the composition and function of mating-type loci in bipolar basidiomycetes. In a landmark study, Bakkeren and Kronstad (2) discovered that in bipolar fungus, Ustilago hordei, the A-and B-mating-type loci were fused into one nonrecombining mating-type region with two alleles. However, subsequent studies revealed that although both A-and B-mating-type homologs are found in bipolar mushrooms, they are present on different chromosomes, and only the A-mating-type homologs are related to mating compatibility (1, 16).Although Ph...

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A -Mating-Type Gene Expression Can Drive Clamp Formation in the Bipolar Mushroom Pholiota microspora (Pholiota nameko)

Mukaiyama

Tachikawa

et al. 2010

Eukaryot Cell

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“…The tetrapolar mating system consists of two sets of mating type genes. The A mating type genes encode homeodomain transcription factors that are assumed to directly regulate gene expression (39,67). A multiallelic pheromone/receptor system is encoded by the B genes.…”

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Ras GTPase-Activating Protein Gap1 of the Homobasidiomycete Schizophyllum commune Regulates Hyphal Growth Orientation and Sexual Development

et al. 2006

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The white rot fungus Schizophyllum commune is used for the analysis of mating and sexual development in homobasidiomycete fungi. In this study, we isolated the gene gap1 encoding a GTPase-activating protein for Ras. Disruption of gap1 should therefore lead to strains accumulating Ras in its activated, GTP-bound state and to constitutive Ras signaling. Haploid ⌬gap1 monokaryons of different mating types did not show alterations in mating behavior in the four different mating interactions possible in fungi expressing a tetrapolar mating type system. Instead, the growth rate in ⌬gap1 monokaryons was reduced by ca. 25% and ca. 50% in homozygous ⌬gap1/⌬gap1 dikaryons. Monokaryons, as well as homozygous dikaryons, carrying the disrupted gap1 alleles exhibited a disorientated growth pattern. Dikaryons showed a strong phenotype during clamp formation since hook cells failed to fuse with the peg beside them. Instead, the dikaryotic character of the hyphae was rescued by fusion of the hooks with nearby developing branches. ⌬gap1/⌬gap1 dikaryons formed increased numbers of fruitbody primordia, whereas the amount of fruitbodies was not raised. Mature fruitbodies formed no or abnormal gills. No production of spores could be observed. The results suggest Ras involvement in growth, clamp formation, and fruitbody development.The homobasidiomycetous white-rot fungus Schizophyllum commune has been used as a model system for the investigation of mating and sexual development for decades since it can be grown from spore to spore through its entire life cycle within 14 days on defined media, and it shows easily distinguished phenotypes for a tetrapolar mating behavior (32, 53). The tetrapolar mating system consists of two sets of mating type genes. The A mating type genes encode homeodomain transcription factors that are assumed to directly regulate gene expression (39,67). A multiallelic pheromone/receptor system is encoded by the B genes. Homology to the yeast pheromone system has led to the expectation that mating is in part controlled by a mitogen-activated protein kinase signal transduction cascade that is activated after stimulation of the G protein-coupled pheromone receptor (20,76,82).A fully compatible mating between two strains of S. commune occurs when both differ in their A and B gene specificities (A B ). The specificity of a locus is defined by a lack of activation of downstream developmental processes after crossing two strains with identical specificities in their mating type genes (20). Several steps of subsequent development can be distinguished. After cell fusion, septal breakdown and fast nuclear migration allow reciprocal nuclear exchange between the two mates. Migrant and resident nuclei pair, and dikaryotic hyphal tips are established. Subsequent conjugate nuclear division is accompanied by formation of clamp connections. Clamp connections are short, backwardly directed branches that fuse with the subapical cell and provide a bypass for one of the nuclei produced during synchronous division of the dikaryon, en...

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“…The monokaryotic and dikaryotic conditions can be distinguished by the presence of clamp connections in dikaryons and their lack in monokaryons. Clamp connections are hookshaped structures involved in equal nuclei sorting to the daughter cells produced by mitosis.Genetic studies carried out in C. cinereus and S. commune have shown that the A incompatibility locus codes for homeodomain-containing transcription factors (2,13,14,18,21,24,27,28). The b mating-type locus of U. maydis is homologous to the A locus and also codes for homeodomain proteins (6, 20).…”

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Relationship between Monokaryotic Growth Rate and Mating Type in the Edible Basidiomycete Pleurotus ostreatus

Larraya

Pérez

Iribarren

et al. 2001

Appl Environ Microbiol

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The edible fungus Pleurotus ostreatus (oyster mushroom) is an industrially produced heterothallic homobasidiomycete whose mating is controlled by a bifactorial tetrapolar genetic system. Two mating loci (matA and matB) control different steps of hyphal fusion, nuclear migration, and nuclear sorting during the onset and progress of the dikaryotic growth. Previous studies have shown that the segregation of the alleles present at the matB locus differs from that expected for a single locus because (i) new nonparental B alleles appeared in the progeny and (ii) there was a distortion in the segregation of the genomic regions close to this mating locus. In this study, we pursued these observations by using a genetic approach based on the identification of molecular markers linked to the matB locus that allowed us to dissect it into two genetically linked subunits (matB␣ and matB␤) and to correlate the presence of specific matB␣ and matA alleles with differences in monokaryotic growth rate. The availability of these molecular markers and the mating type dependence of growth rate in monokaryons can be helpful for marker-assisted selection of fast-growing monokaryons to be used in the construction of dikaryons able to colonize the substrate faster than the competitors responsible for reductions in the industrial yield of this fungus.Incompatibility systems are mechanisms for the creation of variability preventing selfing. The phytopathogenic fungus Ustilago maydis and the mushrooms Coprinus cinereus and Schizophyllum commune have been used as models to study mating incompatibility in basidiomycetes. In these species, mating is controlled by two unlinked multiallelic loci whose independent segregation generates four mating specificities in the progeny of a single individual (these fungi are then called tetrapolar) (for reviews, see references 2, 7, and 11). In tetrapolar basidiomycetes, a single basidiospore produces upon germination a hypha in which all nuclei are identical (homokaryon). Two hyphae with different mating alleles at the two incompatibility loci are able to fuse and give rise to a mycelium in which the two parental nuclei do not fuse throughout vegetative growth. This kind of mycelium is called dikaryotic, and the individual mycelium is called a dikaryon. Vegetative growth is maintained until a set of environmental conditions triggers fruit body formation. Karyogamy occurs within the basidia, and it is immediately followed by meiosis producing four uninucleate spores. The monokaryotic and dikaryotic conditions can be distinguished by the presence of clamp connections in dikaryons and their lack in monokaryons. Clamp connections are hookshaped structures involved in equal nuclei sorting to the daughter cells produced by mitosis.Genetic studies carried out in C. cinereus and S. commune have shown that the A incompatibility locus codes for homeodomain-containing transcription factors (2,13,14,18,21,24,27,28). The b mating-type locus of U. maydis is homologous to the A locus and also codes for homeodomain prote...

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Interaction of the Aα Y and Z Mating-Type Homeodomain Proteins of Schizophyllum commune Detected by the Two-Hybrid System

Cited by 36 publications

References 0 publications

A -Mating-Type Gene Expression Can Drive Clamp Formation in the Bipolar Mushroom Pholiota microspora (Pholiota nameko)

A -Mating-Type Gene Expression Can Drive Clamp Formation in the Bipolar Mushroom Pholiota microspora (Pholiota nameko)

Ras GTPase-Activating Protein Gap1 of the Homobasidiomycete Schizophyllum commune Regulates Hyphal Growth Orientation and Sexual Development

Relationship between Monokaryotic Growth Rate and Mating Type in the Edible Basidiomycete Pleurotus ostreatus

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