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Mycelial form of dimorphic fungus Malassezia species dictates the microbial interaction.

ABSTRACT: Dandruff is one of the most common clinically manifested and studied scalp disorders. It has been associated with both bacteria and fungi. Bacteria and fungi inhabiting the scalp are known to influence each other and manifestation of dandruff. Fungal and bacterial isolates from scalp epithelial flakes (dandruff) were identified by rDNA sequencing. Local oils were tested for fungal and bacterial inhibition, interaction and biofilm formation, cell-cell interactions were studied by auto aggregation and surface thermodynamics studies. The isolates Bacillus sp.C2b1 (MK036745) and Malassezia sp. C2y1 (MK036746) were inhibited by Mahabhrungraj oil. The fungal morphological switch was evident and dependent on nutrition. Cell aggregation studies suggested the interaction of bacteria with yeast (non-pathogenic) phase of the fungus. Bacterial and yeast cells were found to be compatible for biofilm formation. The fungal mycelial surfaces were found to be conducive for interaction with both bacterial cells and yeast forms. The results here indicate the significance of mycelial phase of scalp-isolated fungus in interaction with the bacterial surfaces and also with self-yeast phase surface. This is the first report of the interaction between scalp-isolated microorganisms with respect to their surface interaction capabilities.

SUBMITTER: Patel CD

PROVIDER: S-EPMC6646641 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Mycelial form of dimorphic fungus <i>Malassezia</i> species dictates the microbial interaction.

Patel Charmi D CD Markande Anoop R AR

Indian journal of microbiology 20190320 3

Dandruff is one of the most common clinically manifested and studied scalp disorders. It has been associated with both bacteria and fungi. Bacteria and fungi inhabiting the scalp are known to influence each other and manifestation of dandruff. Fungal and bacterial isolates from scalp epithelial flakes (dandruff) were identified by rDNA sequencing. Local oils were tested for fungal and bacterial inhibition, interaction and biofilm formation, cell-cell interactions were studied by auto aggregation ...[more]

PMID: 31388203

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Project description:BACKGROUND: Penicillium marneffei is the most important thermal dimorphic fungus causing systemic mycosis in China and Southeast Asia. While miRNAs are increasingly recognized for their roles in post-transcriptional regulation of gene expression in animals and plants, miRNAs in fungi were less well studied and their potential roles in fungal dimorphism were largely unknown. Based on P. marneffei genome sequence, we hypothesize that miRNA-like RNAs (milRNAs) may be expressed in the dimorphic fungus. METHODOLOGY/PRINCIPAL FINDINGS: We attempted to identify milRNAs in P. marneffei in both mycelial and yeast phase using high-throughput sequencing technology. Small RNAs were more abundantly expressed in mycelial than yeast phase. Sequence analysis revealed 24 potential milRNA candidates, including 17 candidates in mycelial and seven in yeast phase. Two genes, dcl-1 and dcl-2, encoding putative Dicer-like proteins and the gene, qde-2, encoding Argonaute-like protein, were identified in P. marneffei. Phylogenetic analysis showed that dcl-2 of P. marneffei was more closely related to the homologues in other thermal dimorphic pathogenic fungi than to Penicillium chrysogenum and Aspergillus spp., suggesting the co-evolution of dcl-2 among the thermal dimorphic fungi. Moreover, dcl-2 demonstrated higher mRNA expression levels in mycelial than yeast phase by 7 folds (P<0.001). Northern blot analysis confirmed the expression of two milRNAs, PM-milR-M1 and PM-milR-M2, only in mycelial phase. Using dcl-1(KO), dcl-2(KO), dcl(DKO) and qde-2(KO) deletion mutants, we showed that the biogenesis of both milRNAs were dependent on dcl-2 but not dcl-1 or qde-2. The mRNA expression levels of three predicted targets of PM-milR-M1 were upregulated in knockdown strain PM-milR-M1 (KD), supporting regulatory function of milRNAs. CONCLUSIONS/SIGNIFICANCE: Our findings provided the first evidence for differential expression of milRNAs in different growth phases of thermal dimorphic fungi and shed light on the evolution of fungal proteins involved in milRNA biogenesis and possible role of post-transcriptional control in governing thermal dimorphism.

Project description:BackgroundPenicillium marneffei is a pathogenic fungus that afflicts immunocompromised individuals having lived or traveled in Southeast Asia. This species is unique in that it is the only dimorphic member of the genus. Dimorphism results from a process, termed phase transition, which is regulated by temperature of incubation. At room temperature, the fungus grows filamentously (mould phase), but at body temperature (37 degrees C), a uninucleate yeast form develops that reproduces by fission. Formation of the yeast phase appears to be a requisite for pathogenicity. To date, no genes have been identified in P. marneffei that strictly induce mould-to-yeast phase conversion. In an effort to help identify potential gene products associated with morphogenesis, protein profiles were generated from the yeast and mould phases of P. marneffei.ResultsWhole cell proteins from the early stages of mould and yeast development in P. marneffei were resolved by two-dimensional gel electrophoresis. Selected proteins were recovered and sequenced by capillary-liquid chromatography-nanospray tandem mass spectrometry. Putative identifications were derived by searching available databases for homologous fungal sequences. Proteins found common to both mould and yeast phases included the signal transduction proteins cyclophilin and a RACK1-like ortholog, as well as those related to general metabolism, energy production, and protection from oxygen radicals. Many of the mould-specific proteins identified possessed similar functions. By comparison, proteins exhibiting increased expression during development of the parasitic yeast phase comprised those involved in heat-shock responses, general metabolism, and cell-wall biosynthesis, as well as a small GTPase that regulates nuclear membrane transport and mitotic processes in fungi. The cognate gene encoding the latter protein, designated RanA, was subsequently cloned and characterized. The P. marneffei RanA protein sequence, which contained the signature motif of Ran-GTPases, exhibited 90% homology to homologous Aspergillus proteins.ConclusionThis study clearly demonstrates the utility of proteomic approaches to studying dimorphism in P. marneffei. Moreover, this strategy complements and extends current genetic methodologies directed towards understanding the molecular mechanisms of phase transition. Finally, the documented increased levels of RanA expression suggest that cellular development in this fungus involves additional signaling mechanisms than have been previously described in P. marneffei.

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Mycelial form of dimorphic fungus Malassezia species dictates the microbial interaction.

Publications

Mycelial form of dimorphic fungus <i>Malassezia</i> species dictates the microbial interaction.

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