Cytosolic pH controls fungal MAPK signaling and pathogenicity

In fungi, ambient pH acts as a key regulator of development and virulence. The vascular wilt pathogen Fusarium oxysporum uses host alkalinization to promote infection of plant hosts through activation of the invasive growth mitogen-activated protein kinase (MAPK) Fmk1. The molecular events underlying pH-driven MAPK regulation are unknown. Using the ratiometric GFP-based pH sensor pHluorin, we find that both F. oxysporum and Saccharomyces cerevisiae respond to extracellular alkalinization or acidification with a transitory shift in cytosolic pH (pH c ) and rapid changes in phosphorylation levels of the three fungal MAPKs Fmk1, Mpk1/Slt2 (cell wall integrity) and Hog1 (hyperosmotic stress). Pharmacological inhibition of the essential plasma membrane H+-ATPase Pma1, which leads to pH c acidification, is sufficient to trigger reprogramming of MAPK phosphorylation even in the absence of an extracellular pH shift. Screening of a subset of S. cerevisiae mutants identified the sphingolipid-regulated AGC kinase Ypk1/2 as a key upstream component of pH c -modulated MAPK responses. We further show that acidification of pH c in F. oxysporum leads to an increase of the long chain base (LCB) sphingolipid dihydrosphingosine (dhSph) and that exogenous addition of dhSph activates Mpk1 phosphorylation. Our results reveal a pivotal role of pH c in the regulation of MAPK signaling and suggest new ways to control fungal growth and pathogenicity.