title: A small phospholipase A2-α from castor catalyzes the removal of hydroxy fatty acids from phosphatidylcholine in transgenic Arabidopsis seeds.
abstract: Ricinoleic acid, an industrially useful hydroxy fatty acid (HFA), only accumulates to high levels in the triacylglycerol fraction of castor (Ricinus communis) endosperm, even though it is synthesized on the membrane lipid phosphatidylcholine (PC) from an oleoyl ester. The acyl chains of PC undergo intense remodeling through the process of acyl editing. The identities of the proteins involved in this process, however, are unknown. A phospholipase A2 (PLA2) is thought to be involved in the acyl-editing process. We show here a role for RcsPLA2α in the acyl editing of HFA esterified to PC. RcsPLA2α was identified by its high relative expression in the castor endosperm transcriptome. Coexpression in Arabidopsis (Arabidopsis thaliana) seeds of RcsPLA2α with the castor fatty acid hydroxylase RcFAH12 led to a dramatic decrease in seed HFA content when compared with RcFAH12 expression alone in both PC and the neutral lipid fraction. The low-HFA trait was heritable and gene dosage dependent, with hemizygous lines showing intermediate HFA levels. The low seed HFA levels suggested that RcsPLA2α functions in vivo as a PLA2 with HFA specificity. Activity assays with yeast (Saccharomyces cerevisiae) microsomes showed a high specificity of RcsPLA2α for ricinoleic acid, superior to that of the endogenous Arabidopsis PLA2α. These results point to RcsPLA2α as a phospholipase involved in acyl editing, adapted to specifically removing HFA from membrane lipids in seeds.
authors: Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340 (S.B., J.B.); andAlberta Innovates Phytola Centre, Department of Agricultural, Food, and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5 (G.C., R.J.W.).
authors: Shen
authors: S
authors: Bayon
authors: Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340 (S.B., J.B.); andAlberta Innovates Phytola Centre, Department of Agricultural, Food, and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5 (G.C., R.J.W.).
authors: Guanqun
authors: G
authors: Chen
authors: Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340 (S.B., J.B.); andAlberta Innovates Phytola Centre, Department of Agricultural, Food, and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5 (G.C., R.J.W.).
authors: Randall J
authors: RJ
authors: Weselake
authors: Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340 (S.B., J.B.); andAlberta Innovates Phytola Centre, Department of Agricultural, Food, and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5 (G.C., R.J.W.) jab@wsu.edu.
authors: John
authors: J
authors: Browse
mesh_terms: D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002367:Ricinus communis / Q000201:enzymology* / Q000235:genetics; D005227:Fatty Acids / Q000378:metabolism*; D054513:Group IV Phospholipases A2 / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010713:Phosphatidylcholines / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011133:Polysaccharide-Lyases / Q000235:genetics / Q000378:metabolism; D012277:Ricinoleic Acids / Q000378:metabolism; D012639:Seeds / Q000201:enzymology / Q000235:genetics; D017422:Sequence Analysis, DNA; D014280:Triglycerides / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D005227:Fatty Acids; D010713:Phosphatidylcholines; D010940:Plant Proteins; D012277:Ricinoleic Acids; D014280:Triglycerides; D054513:Group IV Phospholipases A2; D011133:Polysaccharide-Lyases; C513206:PLA2 protein, Arabidopsis; C030521:ricinoleic acid