PMID:24917677

Citation	Dittrich-Domergue, F, Joubès, J, Moreau, P, Lessire, R, Stymne, S and Domergue, F (2014) The Bifunctional Protein TtFARAT from Tetrahymena thermophila Catalyzes the Formation of both Precursors Required to Initiate Ether Lipid Biosynthesis. J. Biol. Chem. 289:21984-94
Abstract	The biosynthesis of ether lipids and wax esters requires as precursors fatty alcohols, which are synthesized by fatty acyl reductases (FARs). The presence of ether glycerolipids as well as branched wax esters has been reported in several free-living ciliate protozoa. In the genome of Tetrahymena thermophila, the only ORF sharing similarities with FARs is fused to an acyltransferase-like domain, whereas, in most other organisms, FARs are monofunctional proteins of similar size and domain structure. Here, we used heterologous expression in plant and yeast to functionally characterize the activities catalyzed by this protozoan protein. Transient expression in tobacco epidermis of a truncated form fused to the green fluorescence protein followed by confocal microscopy analysis suggested peroxisomal localization. In vivo approaches conducted in yeast indicated that the N-terminal FAR-like domain produced both 16:0 and 18:0 fatty alcohols, whereas the C-terminal acyltransferase-like domain was able to rescue the lethal phenotype of the yeast double mutant gat1Δ gat2Δ. Using in vitro approaches, we further demonstrated that this domain is a dihydroxyacetone phosphate acyltransferase that uses preferentially 16:0-coenzyme A as an acyl donor. Finally, coexpression in yeast with the alkyl-dihydroxyacetone phosphate synthase from T. thermophila resulted the detection of various glycerolipids with an ether bond, indicating reconstitution of the ether lipid biosynthetic pathway. Together, these results demonstrate that this FAR-like protein is peroxisomal and bifunctional, providing both substrates required by alkyl-dihydroxyacetone phosphate synthase to initiate ether lipid biosynthesis.
Links	PubMed PMC4139215 Online version:10.1074/jbc.M114.579318
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
TETTS:I7LW01		GO:0016287: glycerone-phosphate O-acyltransferase activity	ECO:0000314:		F		The activity shows a similarity to DHAPAT in figure 4. DHAPAT and GNPAT are used interchangeably for the GO ID's.	complete CACAO 10150
TETTS:I7LW01		GO:0080019: fatty-acyl-CoA reductase (alcohol-forming) activity	ECO:0000314:		F		Figure 2 shows that it is able to produce fatty alcohols just as FAR does.	complete CACAO 10151
TETTS:I7LW01	enables	GO:0016287: glycerone-phosphate O-acyltransferase activity	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
TETTS:I7LW01	enables	GO:0080019: fatty-acyl-CoA reductase (alcohol-forming) activity	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete

References

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