title: Adaptation of the glycerol-3-phosphate dehydrogenase Gpd1 to high salinities in the extremely halotolerant Hortaea werneckii and halophilic Wallemia ichthyophaga.
abstract: We report the first identification and characterisation of the glycerol-3-phosphate dehydrogenase (GPD) genes from extremely halophilic fungi. The black ascomycetous yeast Hortaea werneckii and the non-melanised basidiomycetous fungus Wallemia ichthyophaga inhabit similar hypersaline environments, yet they have two different strategies of haloadaptation through Gpd1-regulated glycerol synthesis. The extremely halotolerant H. werneckii codes for two salt-inducible GPD1 genes that show similar gene transcription regulation and have 98% amino-acid sequence identity between paralogues; however, they have distinct effects when expressed heterologously in Saccharomyces cerevisiae gpd mutants. Only the HwGpd1B isoform complements the function of Gpd in the gpd1 mutant, whereas none of the Gpd1 isoforms can rescue the salt sensitivity of the gpd1gpd2 double mutant. The obligate halophile W. ichthyophaga codes for only one GPD1 orthologue, the transcription of which is less affected by salt when compared to the H. werneckii homologues. Heterologous expression of WiGPD1 in S. cerevisiae recovers halotolerance of the gpd1 and gpd1gpd2 mutant strains, which is probably due to the overall high amino-acid similarity of the Gpd1 protein in W. ichthyophaga and S. cerevisiae. Phylogenetic analysis of amino-acid sequences reveals that the evolutionary origins of all of these three novel enzymes correspond to the phylogeny of the fungal species from which the genes were identified.
authors.affiliation: Institute of Biochemistry, Faculty of Medicine, University of Ljubljana, Vrazov Trg 2, SI-1000 Ljubljana, Slovenia. metka.lenassi@mf.uni-lj.si
authors.forename: Metka
authors.initials: M
authors.lastname: Lenassi
authors.forename: Janja
authors.initials: J
authors.lastname: Zajc
authors.forename: Cene
authors.initials: C
authors.lastname: Gostinčar
authors.forename: Alenka
authors.initials: A
authors.lastname: Gorjan
authors.forename: Nina
authors.initials: N
authors.lastname: Gunde-Cimerman
authors.forename: Ana
authors.initials: A
authors.lastname: Plemenitaš
mesh_terms: D000595:Amino Acid Sequence; D001203:Ascomycota / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D001487:Basidiomycota / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D005993:Glycerolphosphate Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D054712:Salinity; D012623:Seawater / Q000382:microbiology*; D016415:Sequence Alignment; D012965:Sodium Chloride / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins; D012965:Sodium Chloride; D005993:Glycerolphosphate Dehydrogenase