PMID:23537692

Citation	Hagström, AK, Walther, A, Wendland, J and Löfstedt, C (2013) Subcellular localization of the fatty acyl reductase involved in pheromone biosynthesis in the tobacco budworm, Heliothis virescens (Noctuidae: Lepidoptera). Insect Biochem. Mol. Biol. 43:510-21
Abstract	Sex pheromone components are produced in specialized glands of female moths via well-characterized biosynthetic pathways, where a Fatty Acyl Reductase (FAR) is often essential for producing the specific ratio of the different pheromone components. The subcellular localization and membrane topology of FARs is important for understanding how pheromones are synthesized and exported to the exterior for release. We investigated the subcellular localization of HvFAR from the noctuid moth Heliothis virescens by producing recombinant fusion proteins with green fluorescent protein (GFP) in yeast. A C-terminally tagged construct was localized to the endoplasmic reticulum (ER) and retained full reductive activity on a broad range of saturated and unsaturated fatty acyl precursors. In contrast, an N-terminally-tagged construct was poorly expressed in the cytoplasm and was not enzymatically active, indicating that HvFAR requires a free N-terminal for both proper targeting and catalytic activity. A series of truncations of the N-and C-termini of HvFAR was conducted based on in silico-predicted hydrophobic domains and transmembrane regions. The N-terminally truncated protein was found in the cytoplasm and did not retain activity, emphasizing the importance of the N-terminal for FAR function. In addition, the orientation in the membrane of the C-terminus-tagged HvFAR-GFP construct was analyzed using a fluorescence protease protection (FPP) assay, implying that the C-terminal of HvFAR is orientated towards the cytoplasm. These results, together with previous data on the localization of desaturases, confirm the importance of the ER as a subcellular site of pheromone production.
Links	PubMed Online version:10.1016/j.ibmb.2013.03.006
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HELVI:D2SNU9		GO:0005783: endoplasmic reticulum	ECO:0000314:		C		Figures 3A and 4.	complete CACAO 8252
HELVI:D2SNU9		GO:0080019: fatty-acyl-CoA reductase (alcohol-forming) activity	ECO:0000314:		F		Figure S2.	complete CACAO 8253
HELVI:D2SNU9	part_of	GO:0005783: endoplasmic reticulum	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HELVI:D2SNU9	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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