Functionality of the <em>Paracoccidioides</em> Mating α-Pheromone-Receptor System
Jéssica A. Gomes-Rezende
Ana G. Gomes-Alves
João F. Menino
Marco A. Coelho
Paula Ludovico
Paula Gonçalves
Mark H. J. Sturme
Fernando Rodrigues
10.1371/journal.pone.0047033
https://plos.figshare.com/articles/dataset/Functionality_of_the_Paracoccidioides_Mating_Pheromone_Receptor_System/118986
<div><p>Recent evidence suggests that <em>Paracoccidioides</em> species have the potential to undergo sexual reproduction, although no sexual cycle has been identified either in nature or under laboratory conditions. In the present work we detected low expression levels of the heterothallic <em>MAT</em> loci genes <em>MAT1-1</em> and <em>MAT1-2</em>, the α-pheromone (<em>PBα</em>) gene, and the α- and <b>a</b>-pheromone receptor (<em>PREB</em> and <em>PREA</em>) genes in yeast and mycelia forms of several <em>Paracoccidioides</em> isolates. None of the genes were expressed in a mating type dependent manner. Stimulation of <em>P. brasiliensis MAT1-2</em> strains with the synthetic α-pheromone peptide failed to elicit transcriptional activation of <em>MAT1-2, PREB</em> or <em>STE12</em>, suggesting that the strains tested are insensitive to α-pheromone. In order to further evaluate the biological functionality of the pair α-pheromone and its receptor, we took advantage of the heterologous expression of these <em>Paracoccidioides</em> genes in the corresponding <em>S. cerevisiae</em> null mutants. We show that <em>S. cerevisiae</em> strains heterologously expressing <em>PREB</em> respond to Pbα pheromone either isolated from <em>Paracoccidioides</em> culture supernatants or in its synthetic form, both by shmoo formation and by growth and cell cycle arrests. This allowed us to conclude that <em>Paracoccidioides</em> species secrete an active α-pheromone into the culture medium that is able to activate its cognate receptor. Moreover, expression of <em>PREB</em> or <em>PBα</em> in the corresponding null mutants of <em>S. cerevisiae</em> restored mating in these non-fertile strains. Taken together, our data demonstrate pheromone signaling activation by the <em>Paracoccidioides</em> α-pheromone through its receptor in this yeast model, which provides novel evidence for the existence of a functional mating signaling system in <em>Paracoccidioides</em>.</p> </div>
2012-10-04 02:29:46
functionality
mating