Genetic and biochemical studies in yeast reveal that the cotton fibre-specific GhCER6 gene functions in fatty acid elongation.
3-ketoacyl-CoA synthase catalyses the initial condensation reaction during fatty acid elongation using malonyl-CoA and long-chain acyl-CoA as substrates. Previously, it was reported that several genes encoding putative cotton 3-ketoacyl-CoA synthases were significantly up-regulated during early cotton fibre development. In this study, GhCER6 cDNA that contains an open reading frame of 1479 bp, encoding a protein of 492 amino acid residues homologous to the Arabidopsis condensing enzyme CER6, was isolated and cloned. In situ hybridization results demonstrated that GhCER6 mRNA was detected only in the elongating wild-type cotton fibre cells. When GhCER6 was transformed to the Saccharomyces cerevisiae elo3 deletion mutation strain that was deficient in the production of 26-carbon fatty acids and displayed a very slow-growth phenotype, the mutant cells were found to divide similarly compared with those of the wild-type cells. Further, heterologous expression of GhCER6 restored the viability of the S. cerevisiae haploid elo2 and elo3 double-deletion strain. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis showed that GhCER6 was enzymatically active since the yeast elo2 and elo3 double-deletion mutant expressing the cotton gene produced very-long-chain fatty acids that are essential for cell growth. The results suggest that GhCER6 encodes a functional 3-ketoacyl-CoA synthase.
Qin
Yong-Mei
YM
National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, China. qinym@pku.edu.cn
Pujol
François M
FM
Hu
Chun-Yang
CY
Feng
Jian-Xun
JX
Kastaniotis
Alexander J
AJ
Hiltunen
J Kalervo
JK
Zhu
Yu-Xian
YX
D000217:Acyltransferases / Q000737:chemistry / Q000502:physiology; D000595:Amino Acid Sequence; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000502:physiology; D005227:Fatty Acids / Q000378:metabolism*; D005816:Genetic Complementation Test; D003368:Gossypium / Q000201:enzymology / Q000235:genetics*; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D030781:Organisms, Genetically Modified / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D016415:Sequence Alignment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014170:Transformation, Genetic
D029681:Arabidopsis Proteins; D005227:Fatty Acids; D010940:Plant Proteins; D000217:Acyltransferases; C120018:CUT1 protein, Arabidopsis