Characterization of the atypical Ppz/Hal3 phosphatase system from the pathogenic fungusCryptococcus neoformans
| Autor: | David Reverter, Lydia Tabernero, Rocío García-Rodas, Cristina Molero, Haroldo Cesar de Oliveira, Oscar Zaragoza, Chunyi Zhang, Joaquín Ariño |
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| Rok vydání: | 2019 |
| Předmět: |
Models
Molecular Cryptococcus neoformans 0303 health sciences Saccharomyces cerevisiae Proteins biology 030306 microbiology Mutant Phosphatase Saccharomyces cerevisiae Virulence Pathogenic fungus biology.organism_classification Microbiology Fungal Proteins 03 medical and health sciences Phenotype Biochemistry Gene Expression Regulation Fungal Heterotrimeric G protein Cation homeostasis Phosphoprotein Phosphatases Molecular Biology 030304 developmental biology |
| Zdroj: | Molecular Microbiology. 111:898-917 |
| ISSN: | 1365-2958 0950-382X |
| DOI: | 10.1111/mmi.14181 |
| Popis: | Ppz Ser/Thr protein phosphatases (PPases) are found only in fungi and have been proposed as potential antifungal targets. In Saccharomyces cerevisiae Ppz1 (ScPpz1) is involved in regulation of monovalent cation homeostasis. ScPpz1 is inhibited by two regulatory proteins, Hal3 and Vhs3, which have moonlighting properties, contributing to the formation of an unusual heterotrimeric PPC decarboxylase (PPCDC) complex crucial for CoA biosynthesis. Here we report the functional characterization of CnPpz1 (CNAG_03673) and two possible Hal3-like proteins, CnHal3a (CNAG_00909) and CnHal3b (CNAG_07348) from the pathogenic fungus Cryptococcus neoformans. Deletion of CnPpz1 or CnHal3b led to phenotypes unrelated to those observed in the equivalent S. cerevisiae mutants, and the CnHal3b-deficient strain was less virulent. CnPpz1 is a functional PPase and partially replaced endogenous ScPpz1. Both CnHal3a and CnHal3b interact with ScPpz1 and CnPpz1 in vitro but do not inhibit their phosphatase activity. Consistently, when expressed in S. cerevisiae, they poorly reproduced the Ppz1-regulatory properties of ScHal3. In contrast, both proteins were functional monogenic PPCDCs. The CnHal3b isoform was crystallized and, for the first time, the 3D-structure of a fungal PPCDC elucidated. Therefore, our work provides the foundations for understanding the regulation and functional role of the Ppz1-Hal3 system in this important pathogenic fungus. |
| Databáze: | OpenAIRE |
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