Deciphering the Role of PIG1 and DHN-Melanin in Scedosporium apiospermum Conidia

揭示 PIG1 和 DHN-黑色素在尖端赛多孢子霉菌分生孢子中的作用

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作者:Hélène Guegan, Wilfried Poirier, Kevin Ravenel, Sarah Dion, Aymeric Delabarre, Dimitri Desvillechabrol, Xavier Pinson, Odile Sergent, Isabelle Gallais, Jean-Pierre Gangneux, Sandrine Giraud, Amandine Gastebois

Abstract

Scedosporium apiospermum is a saprophytic filamentous fungus involved in human infections, of which the virulence factors that contribute to pathogenesis are still poorly characterized. In particular, little is known about the specific role of dihydroxynaphtalene (DHN)-melanin, located on the external layer of the conidia cell wall. We previously identified a transcription factor, PIG1, which may be involved in DHN-melanin biosynthesis. To elucidate the role of PIG1 and DHN-melanin in S. apiospermum, a CRISPR-Cas9-mediated PIG1 deletion was carried out from two parental strains to evaluate its impact on melanin biosynthesis, conidia cell-wall assembly, and resistance to stress, including the ability to survive macrophage engulfment. ΔPIG1 mutants did not produce melanin and showed a disorganized and thinner cell wall, resulting in a lower survival rate when exposed to oxidizing conditions, or high temperature. The absence of melanin increased the exposure of antigenic patterns on the conidia surface. PIG1 regulates the melanization of S. apiospermum conidia, and is involved in the survival to environmental injuries and to the host immune response, that might participate in virulence. Moreover, a transcriptomic analysis was performed to explain the observed aberrant septate conidia morphology and found differentially expressed genes, underlining the pleiotropic function of PIG1.

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