Abstract

MicroRNAs are molecules involved in post-transcriptional gene regulation. In pathogenic fungi, microRNAs have been described at different morphological stages by regulating targets involved in processes such as morphogenesis and energy production. Members of the Paracoccidioides complex are the main etiological agents of a systemic mycosis in Latin America. Fungi of the Paracoccidioides complex present a wide range of plasticity to colonize different niches. In response to environmental changes these fungi undergo a morphological switch, remodel their cellular metabolism and modulate structural cell wall components. However, the underlying mechanisms regulating the gene expression is not well understood. By using high performance sequencing and bioinformatics analyses, this work characterizes microRNAs produced by Paracoccidioides brasiliensis. Here, we demonstrated that the transcript encoding proteins involved in microRNA biogenesis were differentially expressed in each morphological stage. In addition, 49 microRNAs were identified in cDNA libraries with 44 differentially regulated among the libraries. Sixteen microRNAs were differentially regulated in comparison to the mycelium in the mycelium-to-yeast transition phase. The yeast parasitic phase revealed a complete remodeling of the expression of these small RNAs. Analyses of targets of the induced microRNAs, from the different libraries, revealed that these molecules may potentially regulate in the cell wall, by repressing genes involved in the synthesis and degradation of glucans and chitin. Furthermore, mRNAs involved in cellular metabolism and development were predicted to be regulated by microRNAs. Therefore, this work describes a putative post transcriptional regulation, mediated by microRNAs in P. brasiliensis and its influence on the adaptive processes of thermal dimorphic fungus.

Highlights

  • Small non-coding RNAs play an essential regulatory role in biological systems, without being translated into proteins

  • The procedure for extracting RNAs of the different phases of P. brasiliensis was carried out with the purpose of identifying microRNAs produced by this pathogen

  • The transcript coding for Dcr-1 was up-regulated during the transition and in the yeast phase, while Dcr-2 was downregulated during the mycelium-to-yeast transition

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Summary

Introduction

Small non-coding RNAs play an essential regulatory role in biological systems, without being translated into proteins. Of these RNAs, microRNAs are small RNAs ranging in size from 21 to 24 nt and their action is to post-transcriptionally regulate the expression of target genes involved in different processes such as cell proliferation, tumorigenesis, and infection (Bartel, 2004). Expression of microRNAs is regulated under a wide range of different conditions, such as changes in temperature (Bai et al, 2015) and growth in different morphologies (Zhou et al, 2012; Lau et al, 2013). MicroRNA produced by Metarhizium anisopliae, target mRNAs encoding proteins involved in sporulation and may influence the process of conidia formation (Zhou et al, 2012)

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