Functional characterization of a plastidial omega-3 desaturase from sunflower ( Helianthus annuus) in cyanobacteria

• Published in: Plant physiology and biochemistry 2006-10, Vol.44 (10), p.517-525
• Main Authors: Venegas-Calerón, M., Muro-Pastor, A.M., Garcés, R., Martínez-Force, E.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Biological and medical sciences; Chloroplast; Cloning, Molecular; Cyanobacteria; Cyanobacteria - genetics; Cyanobacteria - metabolism; Cyanophyta; DNA, Complementary; Enzymes; Fad7; Fatty acid; Fatty Acid Desaturases - genetics; Fatty Acid Desaturases - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Plant; Helianthus; Helianthus - enzymology; Helianthus - genetics; Helianthus annuus; Helianthus annuus; Heterologous expression; Metabolism; Molecular Sequence Data; Omega-3 desaturase; Phylogeny; Plant physiology and development; Plastids - enzymology; Synechocystis; Synechocystis - genetics; Synechocystis - metabolism
• ISSN: 0981-9428; 1873-2690
• DOI: 10.1016/j.plaphy.2006.09.005

Fatty acid desaturases (FAD) play an important role in plant lipid metabolism and they can be found in several subcellular compartments such as the plastids and endoplasmic reticulum. Lipids are critical components of the cell membrane and, as a consequence, they are fundamental for the proper growth and development of all living organisms. We have used sequences from the conserved regions of known omega-3-desaturases to design degenerated oligonucleotides and clone a cDNA encoding a plastidial omega-3 desaturase from sunflower ( HaFAD7). From its presumed full-length sequence, we predict that Hafad7 encodes a protein of 443 amino acids with a molecular mass of 50.8 kDa, and that it contains a putative chloroplast transit peptide of 51 amino acids. The predicted hydrophobicity of the protein identifies four potential membrane-spanning regions and, according to the TargetP algorithm, the protein should be targeted to the plastid/chloroplast membrane. RT-PCR analysis of its expression shows the transcript is preferentially expressed in photosynthetically active tissues. Heterologous expression of this protein in the unicellular cyanobacterium Synechocystis sp. PCC 6803 confirmed that the protein produced from this cDNA has omega-3 desaturase activity.