caa a22 4500 445318112 CHVBK 20180317142654.0 cr unu---uuuuu 170323e20110201xx s 000 0 eng 10.1007/s10126-010-9264-4 doi (NATIONALLICENCE)springer-10.1007/s10126-010-9264-4 Cloning, Tissue Expression Analysis, and Functional Characterization of Two Δ6-Desaturase Variants of Sea Bass ( Dicentrarchus labrax L.) [Elektronische Daten] [Ester Santigosa, Florian Geay, Thierry Tonon, Herve Le Delliou, Heiner Kuhl, Richard Reinhardt, Laurent Corcos, Chantal Cahu, José Zambonino-Infante, David Mazurais] Fish are the main source of the n-3 highly unsaturated fatty acids, which are crucial for human health. Their synthesis from C18 precursors is mediated by desaturases and elongases, but the activity of these enzymes has not been conclusively established in marine fish species. This study reports the cloning, tissue expression, and functional characterization of a sea bass (Dicentrarchus labrax L.) Δ6-desaturase and one of its splicing variants. Two cDNAs with open reading frames of 1,346 and 1,354bp were cloned and named D6D and D6D-V, respectively. Both deduced protein sequences (445 and 387 amino acids, respectively) contained two transmembrane regions and the N-terminal cytochrome b5 domain with the HPGG motif characteristic of microsomal desaturases. D6D presents three histidine-rich regions, whereas in D6D-V, an insertion of eight nucleotides in the boundaries of exons 10 and 11 modified the third histidine-rich domain and led to insertion of a premature STOP codon, resulting in a shorter predicted protein. Quantitative real-time polymerase chain reaction assay of gene expression showed that D6D was highly expressed in the brain and intestine, and to a lesser extent, in muscle and liver; meanwhile, D6D-V was expressed in all tissues tested, but at level at least 200-fold lower than D6D. Functional analysis in yeast showed that sea bass D6D encodes a fully functional Δ6-desaturase with no residual Δ5-desaturase activity. This desaturase does not exhibit a clear preference for n-3 versus n-6 C18 substrates. Interestingly, D6D-V is a nonfunctional protein, suggesting that the C-terminal end is indispensable for protein activity. Springer Science+Business Media, LLC, 2010 Sea bass ( Dicentrarchus labrax ) nationallicence Desaturase nationallicence HUFA biosynthesis nationallicence Fish nationallicence EPA nationallicence Santigosa Ester Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut Geay Florian Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut Tonon Thierry UPMC Univ Paris 6, UMR 7139 Végétaux marins et biomolécules, Station Biologique, 29682, Roscoff, France aut Le Delliou Herve Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut Kuhl Heiner Max-Planck-Institute Molecular Genetics, Ihnestr. 63-73, 14195, Berlin-Dahlem, Germany aut Reinhardt Richard Max-Planck-Institute Molecular Genetics, Ihnestr. 63-73, 14195, Berlin-Dahlem, Germany aut Corcos Laurent Université de Brest, INSERM, U613, ECLA, 29200, Brest, France aut Cahu Chantal Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut Zambonino-Infante José Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut Mazurais David Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut Marine Biotechnology Springer-Verlag 13/1(2011-02-01), 22-31 1436-2228 13:1<22 2011 13 10126 https://doi.org/10.1007/s10126-010-9264-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10126-010-9264-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Santigosa Ester Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut NATIONALLICENCE 700 1- Geay Florian Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut NATIONALLICENCE 700 1- Tonon Thierry UPMC Univ Paris 6, UMR 7139 Végétaux marins et biomolécules, Station Biologique, 29682, Roscoff, France aut NATIONALLICENCE 700 1- Le Delliou Herve Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut NATIONALLICENCE 700 1- Kuhl Heiner Max-Planck-Institute Molecular Genetics, Ihnestr. 63-73, 14195, Berlin-Dahlem, Germany aut NATIONALLICENCE 700 1- Reinhardt Richard Max-Planck-Institute Molecular Genetics, Ihnestr. 63-73, 14195, Berlin-Dahlem, Germany aut NATIONALLICENCE 700 1- Corcos Laurent Université de Brest, INSERM, U613, ECLA, 29200, Brest, France aut NATIONALLICENCE 700 1- Cahu Chantal Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut NATIONALLICENCE 700 1- Zambonino-Infante José Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut NATIONALLICENCE 700 1- Mazurais David Ifremer Marine Fish Nutrition Team, Nutrition Aquaculture and Genomics Research Unit, UMR 1067, Ifremer, Technopole Brest-Iroise, BP 70, 29280, Plouzané, France aut NATIONALLICENCE 773 0- Marine Biotechnology Springer-Verlag 13/1(2011-02-01), 22-31 1436-2228 13:1<22 2011 13 10126 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer