|
|
|
Long Chain Acyl-Coenzyme A Synthetase 4 (BnLACS4) Gene from Brassica napus Enhances the Yeast Lipid Contents |
TAN Xiao-li, ZHENG Xiang-feng, ZHANG Zhi-yan, WANG Zheng, XIA Heng-chuan, LU Changming, GU Shou-lai |
1.Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, P.R.China
2.Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, P.R.China |
|
|
摘要 Long-chain acyl-Coenzyme A (CoA) synthetases (LACSs) catalyze the formation of long-chain acyl-CoA, and play important roles in fatty acid metabolism including phospholipids, triacylglycerol (TAG) biosynthesis and fatty acid β-oxidation. Here, we report the characterization of a LACS gene from Brassica napus. It is highly homologous to Arabidopsis LACS4 and thus designated as BnLACS4. The cloned gene BnLACS4 could complement a LACS-deficient yeast strain YB525. It is mainly expressed in flowers and developing seeds where lipid biosynthesis is at high rate in Brassica napus. When transiently expressed in tobacco leaves, BnLACS4 is localized in endoplasmic reticulum (ER), the common site for eukaryotic pathway of lipid biosynthesis. Expression of BnLACS4 in the yeast strain pep4 increased its lipid content. Taken together, our results suggest that BnLACS4 may be involved in lipid biosynthesis in B. napus.
Abstract Long-chain acyl-Coenzyme A (CoA) synthetases (LACSs) catalyze the formation of long-chain acyl-CoA, and play important roles in fatty acid metabolism including phospholipids, triacylglycerol (TAG) biosynthesis and fatty acid β-oxidation. Here, we report the characterization of a LACS gene from Brassica napus. It is highly homologous to Arabidopsis LACS4 and thus designated as BnLACS4. The cloned gene BnLACS4 could complement a LACS-deficient yeast strain YB525. It is mainly expressed in flowers and developing seeds where lipid biosynthesis is at high rate in Brassica napus. When transiently expressed in tobacco leaves, BnLACS4 is localized in endoplasmic reticulum (ER), the common site for eukaryotic pathway of lipid biosynthesis. Expression of BnLACS4 in the yeast strain pep4 increased its lipid content. Taken together, our results suggest that BnLACS4 may be involved in lipid biosynthesis in B. napus.
|
Received: 06 January 2013
Accepted:
|
Fund: This work was supported by the Natural Science Foundation of China (31271760) and the Opening Funds of the Key Laboratory of Biology and Genetic Improvement of Oilcrops, Ministry of Agriculture, China. |
Corresponding Authors:
TAN Xiao-li, Fax: +86-511-88791923, E-mail: xltan@ujs.edu.cn; GU Shou-lai, E-mail: gushoulai1987@126.com
E-mail: xltan@ujs.edu.cn; GU Shou-lai, E-mail: gushoulai1987@126.com
|
About author: TAN Xiao-li, Fax: +86-511-88791923, E-mail: xltan@ujs.edu.cn |
Cite this article:
TAN Xiao-li, ZHENG Xiang-feng, ZHANG Zhi-yan, WANG Zheng, XIA Heng-chuan, LU Changming, GU Shou-lai.
2014.
Long Chain Acyl-Coenzyme A Synthetase 4 (BnLACS4) Gene from Brassica napus Enhances the Yeast Lipid Contents. Journal of Integrative Agriculture, 13(1): 54-62.
|
Bach L, Michaelson LV, Haslam R, Bellec Y, Gissot L,Marion J, da Costa M, Boutin JP, Miquel M, Tellier F,et al. 2008. The very-long-chain hydroxy fatty acyl-CoAdehydratase PASTICCINO2 is essential and limitingfor plant development. Proceedings of the National Academy of Sciences of the United States of America,38, 14727-14731Bailey T L, Elkan C. 1994. Fitting a mixture model by expectation maximization to discover motifs inbiopolymers. Proceedings of International Conference on Intelligent Systems for Molecular Biology, 2, 28-36Beaudoin F, Wu X, Li F, Haslam R P, Markham J E, Zheng H,Napier J A, Kunst L. 2009. Functional characterizationof the Arabidopsis beta-ketoacyl-coenzyme A reductasecandidates of the fatty acid elongase. Plant Physiology, 3,1174-1191Black P N, DiRusso C C, Metzger A K, Heimert T L. 1992.Cloning, sequencing, and expression of the fadD gene ofEscherichia coli encoding acyl coenzyme A synthetase.The Journal of Biological Chemistry, 35, 25513-25520Chen S, Songkumarn P, Liu J, Wang G L. 2009. A versatilezero background T-vector system for gene cloning andfunctional genomics. Plant Physiology, 3, 1111-1121Choi J Y, Martin C E. 1999. The Saccharomyces cerevisiaeFAT1 gene encodes an acyl-CoA synthetase that is required for maintenance of very long chain fatty acidlevels. The Journal of Biological Chemistry, 8, 4671-4683Dyer J M, Stymne S, Green A G, Carlsson A S. 2008. High-value oils from plants. The Plant Journal, 4, 640-655Faergeman N J, Black P N, Zhao X D, Knudsen J, DiRussoC C. 2001. The Acyl-CoA synthetases encoded withinFAA1 and FAA4 in Saccharomyces cerevisiae functionas components of the fatty acid transport system linkingimport, activation, and intracellular utilization. TheJournal of Biological Chemistry, 40, 37051-37059Faergeman N J, Knudsen J. 1997. Role of long-chain fattyacyl-CoA esters in the regulation of metabolism and in cell signalling. The Biochemical Journal, 323, 1-12Fulda M, Heinz E,Wolter F P. 1997. Brassica napus cDNAsencoding fatty acyl-CoA synthetase. Plant Molecular Biology, 5, 911-922Fulda M, Schnurr J, Abbadi A, Heinz E, Browse J. 2004.Peroxisomal Acyl-CoA synthetase activity is essential for seedling development in Arabidopsis thaliana. ThePlant Cell, 2, 394-405Gietz R D, Schiestl R H. 2007. Frozen competent yeast cellsthat can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method. Nature Protocols, 1,1-4Groot P H, Scholte H R, Hulsmann W C. 1976. Fattyacid activation: specificity, localization, and function.Advances in Lipid Research, 14, 75-126Harwood J L. 1996. Recent advances in the biosynthesis ofplant fatty acids. Biochimica et Biophysica Acta, 1301,7-56Hayashi H, de Bellis L, Hayashi Y, Nito K, Kato A, HayashiM, Hara-Nishimura I, Nishimura M. 2002. Molecular characterization of an Arabidopsis acyl-coenzyme a synthetase localized on glyoxysomal membranes. PlantPhysiology, 4, 2019-2026Igal R A, Wang P, Coleman R A. 1997. Triacsin C blocksde novo synthesis of glycerolipids and cholesterol esters but not recycling of fatty acid into phospholipid:evidence for functionally separate pools of acyl-CoA.The Biochemical Journal, 324, 529-534Jessen D, Olbrich A, Knufer J, Kruger A, Hoppert M, PolleA, Fulda M. 2011. Combined activity of LACS1 andLACS4 is required for proper pollen coat formation inArabidopsis. The Plant Journal, 68, 715-726Karimi M, Inze D, Depicker A. 2002. GATEWAY vectorsfor Agrobacterium-mediated plant transformation.Trends in Plant Science, 5, 193-195Li J, Zhao-Hui C, Batoux M, Nekrasov V, Roux M, Chinchilla D, Zipfel C, Jones J D. 2009. Specific ER quality control components required for biogenesis of the plant innate immune receptor EFR. Proceedings of the National Academy of Sciences of the United States ofAmerica, 37, 15973-15978Lu S, Song T, Kosma D K, Parsons E P, Rowland O,Jenks M A. 2009. Arabidopsis CER8 encodes LONG-CHAIN ACYL-COA SYNTHETASE 1 (LACS1) that has overlapping functions with LACS2 in plant wax andcutin synthesis. The Plant Journal, 59, 553-564Mashek D G, Bornfeldt K E, Coleman R A, Berger J,Bernlohr D A, Black P, DiRusso C C, Farber S A, GuoW, Hashimoto N, et al. 2004. Revised nomenclature forthe mammalian long-chain acyl-CoA synthetase gene family. The Journal of Lipid Research, 10, 1958-1961Ohlrogge J, Browse J. 1995. Lipid biosynthesis. The PlantCell, 7, 957-970Ohlrogge J B. 1994. Design of new plant products: engineering of fatty acid metabolism. Plant Physiology, 3,821-826Pongdontri P, Hills M. 2001. Characterization of a novel plant acyl-coA synthetase that is expressed in lipogenictissues of Brassica napus L. Plant Molecular Biology, 6,717-726Saitou N, Nei M. 1987. The neighbor-joining method: a newmethod for reconstructing phylogenetic trees. MolecularBiology and Evolution, 4, 406-425Schnurr J, Shockey J, Browse J. 2004. The acyl-CoA synthetase encoded by LACS2 is essential for normal cuticle development in Arabidopsis. The Plant Cell, 3,629-642Schnurr J A, Shockey J M, de Boer G J, Browse J A. 2002.Fatty acid export from the chloroplast. Molecular characterization of a major plastidial acyl-coenzyme A synthetase from Arabidopsis. Plant Physiology, 4, 1700-1709Shockey J M, Fulda M S,Browse J A. 2002. Arabidopsiscontains nine long-chain acyl-coenzyme a synthetase genes that participate in fatty acid and glycerolipidmetabolism. Plant Physiology, 4, 1710-1722Slabas A R, Fawcett T. 1992. The biochemistry andmolecular biology of plant lipid biosynthesis. PlantMolecular Biology, 1, 169-191Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 8,1596-1599Tang P Z, Tsai-Morris C H, Dufau M L. 2001. Cloningand characterization of a hormonally regulated rat longchain acyl-CoA synthetase. Proceedings of the NationalAcademy of Sciences of the United States of America,12, 6581-6586Thakur M S, Prapulla S G, Karanth N G. 1989. Estimationof intracellular lipids by the measurement of absorbanceof yeast cells stained with sudan black B. Enzyme andMicrobial Technology, 11, 252-254Thelen J J, Ohlrogge J B. 2002. Metabolic engineering of fatty acid biosynthesis in plants. Metabolic Engineering,1, 12-21Tonon T, Qing R, Harvey D, Li Y, Larson T R, Graham IA. 2005. Identification of a long-chain polyunsaturatedfatty acid acyl-coenzyme A synthetase from the diatomThalassiosira pseudonana. Plant Physiology, 1, 402-408Trenkamp S, Martin W, Tietjen K. 2004. Specific anddifferential inhibition of very-long-chain fatty acidelongases from Arabidopsis thaliana by different herbicides. Proceedings of the National Academy of Sciences of the United States of America, 32, 11903-11908Wang X L, Li X B. 2009. The GhACS1 gene encodes an acyl-CoA synthetase which is essential for normal microsporogenesis in early anther development of cotton. The Plant Journal, 3, 473-486Weng H, Molina I, Shockey J, Browse J. 2010. Organ fusioand defective cuticle function in a lacs1 lacs2 double mutant of Arabidopsis. Planta, 5, 1089-1100Wood C C, Petrie J R, Shrestha P, Mansour M P, Nichols P D,Green A G, Singh S P. 2009. A leaf-based assay usinginterchangeable design principles to rapidly assemble multistep recombinant pathways. Plant Biotechnology Journal, 9, 914-924Zhao L, Katavic V, Li F, Haughn G W, Kunst L. 2010.Insertional mutant analysis reveals that long-chainacyl-CoA synthetase 1 (LACS1), but not LACS8,functionally overlaps with LACS9 in Arabidopsis seedoil biosynthesis. The Plant Journal, 6, 1048-1058Zheng H, Rowland O, Kunst L. 2005. Disruptions of the Arabidopsis Enoyl-CoA reductase gene reveal anessential role for very-long-chain fatty acid synthesis incell expansion during plant morphogenesis. The PlantCell, 5, 1467-1481Zubenko G S, Park F J, Jones E W. 1983. Mutations in PEP4 locus of Saccharomyces cerevisiae block final stepin maturation of two vacuolar hydrolases. Proceedingsof the National Academy of Sciences of the United Statesof America, 2, 510-514 |
No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
|
Shared |
|
|
|
|
|
Discussed |
|
|
|
|