Yeast filamentation signaling is connected to a specific substrate translocation mechanism of the Mep2 transceptor

Autoři: Ana Sofia Brito aff001;  Benjamin Neuhäuser aff002;  René Wintjens aff003;  Anna Maria Marini aff001;  Mélanie Boeckstaens aff001
Působiště autorů: Biology of Membrane Transport Laboratory, Molecular Biology Department, Université Libre de Bruxelles, Gosselies, Belgium aff001;  Institute of Crop Science, Nutritional Crop Physiology, University of Hohenheim, Stuttgart, Germany aff002;  Unité Microbiologie, Chimie Bioorganique et Macromoléculaire, Département R3D, Faculté de Pharmacie, Université Libre de Bruxelles, Brussels, Belgium aff003;  Unité Microbiologie, Chimie Bioorganique et Macromoléculaire, Département RD3, Faculté de Pharmacie, Université Libre de Bruxelles, Brussels, Belgium aff003
Vyšlo v časopise: Yeast filamentation signaling is connected to a specific substrate translocation mechanism of the Mep2 transceptor. PLoS Genet 16(2): e32767. doi:10.1371/journal.pgen.1008634
Kategorie: Research Article


The dimorphic transition from the yeast to the filamentous form of growth allows cells to explore their environment for more suitable niches and is often crucial for the virulence of pathogenic fungi. In contrast to their Mep1/3 paralogues, fungal Mep2-type ammonium transport proteins of the conserved Mep-Amt-Rh family have been assigned an additional receptor role required to trigger the filamentation signal in response to ammonium scarcity. Here, genetic, kinetic and structure-function analyses were used to shed light on the poorly characterized signaling role of Saccharomyces cerevisiae Mep2. We show that Mep2 variants lacking the C-terminal tail conserve the ability to induce filamentation, revealing that signaling can proceed in the absence of exclusive binding of a putative partner to the largest cytosolic domain of the protein. Our data support that filamentation signaling requires the conformational changes accompanying substrate translocation through the pore crossing the hydrophobic core of Mep2. pHluorin reporter assays show that the transport activity of Mep2 and of non-signaling Mep1 differently affect yeast cytosolic pH in vivo, and that the unique pore variant Mep2H194E, with apparent uncoupling of transport and signaling functions, acquires increased ability of acidification. Functional characterization in Xenopus oocytes reveals that Mep2 mediates electroneutral substrate translocation while Mep1 performs electrogenic transport. Our findings highlight that the Mep2-dependent filamentation induction is connected to its specific transport mechanism, suggesting a role of pH in signal mediation. Finally, we show that the signaling process is conserved for the Mep2 protein from the human pathogen Candida albicans.

Klíčová slova:

Candida albicans – Histidine – Multiple alignment calculation – Proline – Saccharomyces cerevisiae – Sequence alignment – Sequence databases – Xenopus oocytes


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