title: Trehalose/2-sulfotrehalose biosynthesis and glycine-betaine uptake are widely spread mechanisms for osmoadaptation in the Halobacteriales.
abstract: We investigated the mechanisms of osmoadaptation in the order Halobacteriales, with special emphasis on Haladaptatus paucihalophilus, known for its ability to survive in low salinities. H. paucihalophilus genome contained genes for trehalose synthesis (trehalose-6-phosphate synthase/trehalose-6-phosphatase (OtsAB pathway) and trehalose glycosyl-transferring synthase pathway), as well as for glycine betaine uptake (BCCT family of secondary transporters and QAT family of ABC transporters). H. paucihalophilus cells synthesized and accumulated ∼1.97-3.72 μmol per mg protein of trehalose in a defined medium, with its levels decreasing with increasing salinities. When exogenously supplied, glycine betaine accumulated intracellularly with its levels increasing at higher salinities. RT-PCR analysis strongly suggested that H. paucihalophilus utilizes the OtsAB pathway for trehalose synthesis. Out of 83 Halobacteriales genomes publicly available, genes encoding the OtsAB pathway and glycine betaine BCCT family transporters were identified in 38 and 60 genomes, respectively. Trehalose (or its sulfonated derivative) production and glycine betaine uptake, or lack thereof, were experimentally verified in 17 different Halobacteriales species. Phylogenetic analysis suggested that trehalose synthesis is an ancestral trait within the Halobacteriales, with its absence in specific lineages reflecting the occurrence of gene loss events during Halobacteriales evolution. Analysis of multiple culture-independent survey data sets demonstrated the preference of trehalose-producing genera to saline and low salinity habitats, and the dominance of genera lacking trehalose production capabilities in permanently hypersaline habitats. This study demonstrates that, contrary to current assumptions, compatible solutes production and uptake represent a common mechanism of osmoadaptation within the Halobacteriales.
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA.
authors.forename: Noha H
authors.initials: NH
authors.lastname: Youssef
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA.|3Present address: Department of Biology, Georgia State University, Atlanta, Georgia.
authors.forename: Kristen N
authors.initials: KN
authors.lastname: Savage-Ashlock
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA.|4Present address: Department of Biology, Indiana University, Bloomington IN.
authors.forename: Alexandra L
authors.initials: AL
authors.lastname: McCully
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA.|5Present address: US Department of Agriculture, Agricultural Research Service, Roman L. Hruska Meat Animal Research Center, Clay Center, NE 68933-0166.
authors.forename: Brandon
authors.initials: B
authors.lastname: Luedtke
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA.
authors.forename: Edward I
authors.initials: EI
authors.lastname: Shaw
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA.|Department of Chemistry, Oklahoma State University, Stillwater, OK, USA.
authors.forename: Wouter D
authors.initials: WD
authors.lastname: Hoff
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA. Mostafa@okstate.edu.
authors.forename: Mostafa S
authors.initials: MS
authors.lastname: Elshahed
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D001622:Betaine / Q000378:metabolism*; D017753:Ecosystem; D020869:Gene Expression Profiling; D005964:Glucosyltransferases / Q000378:metabolism; D016931:Halobacteriales / Q000145:classification / Q000235:genetics / Q000502:physiology*; D026901:Membrane Transport Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D054712:Salinity; D012965:Sodium Chloride / Q000378:metabolism; D014199:Trehalose / Q000096:biosynthesis
chemical_list: D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D001622:Betaine; D012965:Sodium Chloride; D014199:Trehalose; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase
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