BackgroundIt is well known that mushrooms produce defense proteins and secondary metabolites against predators and competitors; however, less is known about the correlation between the tissue-specific expression and the target organism (antagonist) specificity of these molecules. In addition, conserved transcriptional circuitries involved in developing sexual organs in fungi are not characterized, despite the growing number of gene expression datasets available from reproductive and vegetative tissue. The aims of this study were: first, to evaluate the tissue specificity of defense gene expression in the model mushroom Coprinopsis cinerea and, second, to assess the degree of conservation in transcriptional regulation during sexual development in basidiomycetes.ResultsIn order to characterize the regulation in the expression of defense loci and the transcriptional circuitries controlling sexual reproduction in basidiomycetes, we sequenced the poly (A)-positive transcriptome of stage 1 primordia and vegetative mycelium of C. cinerea A43mutB43mut. Our data show that many genes encoding predicted and already characterized defense proteins are differentially expressed in these tissues. The predicted specificity of these proteins with regard to target organisms suggests that their expression pattern correlates with the type of antagonists these tissues are confronted with. Accordingly, we show that the stage 1 primordium-specific protein CC1G_11805 is toxic to insects and nematodes. Comparison of our data to analogous data from Laccaria bicolor and Schizophyllum commune revealed that the transcriptional regulation of nearly 70 loci is conserved and probably subjected to stabilizing selection. A Velvet domain-containing protein was found to be up-regulated in all three fungi, providing preliminary evidence of a possible role of the Velvet protein family in sexual development of basidiomycetes. The PBS-soluble proteome of C. cinerea primordia and mycelium was analyzed by shotgun LC-MS. This proteome data confirmed the presence of intracellular defense proteins in primordia.ConclusionsThis study shows that the exposure of different tissues in fungi to different types of antagonists shapes the expression pattern of defense loci in a tissue-specific manner. Furthermore, we identify a transcriptional circuitry conserved among basidiomycetes during fruiting body formation that involves, amongst other transcription factors, the up-regulation of a Velvet domain-containing protein.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-492) contains supplementary material, which is available to authorized users.

Background: Galactosylation of core fucose residues is a peculiar feature of invertebrate N-glycans. Results: Two variants of galactosylated fucose were observed in a Caenorhabditis elegans mutant and in two nematode parasites. Conclusion: Galactosylation of fucose in nematodes is more widespread than previously thought and is affected by processing hexosaminidases. Significance: Galactosylated fucose motifs may present new targets for anthelminthic agents.

The dung of herbivores, the natural habitat of the model mushroom Coprinopsis cinerea, is a nutrient-rich but also very competitive environment for a saprophytic fungus. We showed previously that C. cinerea expresses constitutive, tissue-specific armories against antagonists such as animal predators and bacterial competitors. In order to dissect the inducible armories against such antagonists, we sequenced the poly(A)-positive transcriptome of C. cinerea vegetative mycelium upon challenge with fungivorous and bacterivorous nematodes, Gram-negative and Gram-positive bacteria and mechanical damage. As a response to the fungivorous nematode Aphelenchus avenae, C. cinerea was found to specifically induce the transcription of several genes encoding previously characterized nematotoxic lectins. In addition, a previously not characterized gene encoding a cytoplasmic protein with several predicted Ricin B-fold domains, was found to be strongly upregulated under this condition. Functional analysis of the recombinant protein revealed a high toxicity toward the bacterivorous nematode Caenorhabditis elegans. Challenge of the mycelium with A. avenae also lead to the induction of several genes encoding putative antibacterial proteins. Some of these genes were also induced upon challenge of the mycelium with the bacteria Escherichia coli and Bacillus subtilis. These results suggest that fungi have the ability to induce specific innate defense responses similar to plants and animals.

Our results support the hypothesis that cytoplasmic proteins abundant in fungal fruiting bodies are involved in fungal resistance against predation. The toxicity of these proteins toward stylet-feeding nematodes, which are also capable of feeding on plants, and the abundance of these proteins in edible mushrooms, may open possible avenues for biological crop protection against parasitic nematodes, e.g., by expression of these proteins in crops.

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Current methods for evaluating mycobacterial invasion of target cells pose technical difficulties including a long turn-around time. Thus, new methodologies must be developed that allow rapid and reliable monitoring of host cell invasion. Here, the invasion of A549 cell line by SYBR safe-labeled Mycobacterium tuberculosis (M. tuberculosis) H37Rv was assessed by flow cytometry and was expressed as the percentage of cells infected by M. tuberculosis. Over a third of A549 cells were invaded by M. tuberculosis, two hours after infection. The specificity of the invasion was confirmed in assays using red blood cells as target cells and Escherichia coli as the non-invasive bacterial control. These findings show that invasion of pulmonary epithelial cells by M. tuberculosis can rapidly and quantitatively be assessed with a great sensitivity by flow cytometric detection of SYBR safe-labeled M. tuberculosis.

Tuberculosis (TB) is a major public health problem throughout the world, affecting almost nine million people [1] and causing more than three million deaths per year. An increasing incidence of TB, related to the high risk of developing the disease in immunosuppressed individuals and the increasing proportion of Mycobacterium tuberculosis drug-resistant strains, has contributed to this problem [2,3]. This makes the Keywords high-activity binding peptide; invasion inhibition; Mycobacterium tuberculosishost cell interaction; Rv2560 membrane protein Correspondence M. E. Patarroyo, Carrera 50 # 26-00,

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