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Recent evidence suggests that  Paracoccidioides  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  MAT  loci genes  MAT1-1  and  MAT1-2 , the α-pheromone ( PBα ) gene, and the α- and  a -pheromone receptor ( PREB  and  PREA ) genes in yeast and mycelia forms of several  Paracoccidioides  isolates. None of the genes were expressed in a mating type dependent manner. Stimulation of  P. brasiliensis MAT1-2  strains with the synthetic α-pheromone peptide failed to elicit transcriptional activation of  MAT1-2, PREB  or  STE12 , 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  Paracoccidioides  genes in the corresponding  S. cerevisiae  null mutants. We show that  S. cerevisiae  strains heterologously expressing  PREB  respond to Pbα pheromone either isolated from  Paracoccidioides  culture supernatants or in its synthetic form, both by shmoo formation and by growth and cell cycle arrests. This allowed us to conclude that  Paracoccidioides  species secrete an active α-pheromone into the culture medium that is able to activate its cognate receptor. Moreover, expression of  PREB  or  PBα  in the corresponding null mutants of  S. cerevisiae  restored mating in these non-fertile strains. Taken together, our data demonstrate pheromone signaling activation by the  Paracoccidioides  α-pheromone through its receptor in this yeast model, which provides novel evidence for the existence of a functional mating signaling system in  Paracoccidioides .

Functionality of the Paracoccidioides Mating α-Pheromone-Receptor System