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Journal of Integrative Agriculture  2014, Vol. 13 Issue (1): 54-62    DOI: 10.1016/S2095-3119(13)60583-3
Crop Genetics · Breeding · Germplasm Resources Advanced Online Publication | Current Issue | Archive | Adv Search |
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
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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.
Keywords:  Brasscia napus       LACS       lipid       subcellular localization       yeast  
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, 14727-14731

Bailey 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, 28-36

Beaudoin 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, 3,1174-1191

Black 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, 25513-25520

Chen S, Songkumarn P, Liu J, Wang G L. 2009. A versatilezero background T-vector system for gene cloning andfunctional genomics. Plant Physiology, 3, 1111-1121

Choi 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, 4671-4683

Dyer J M, Stymne S, Green A G, Carlsson A S. 2008. High-value oils from plants. The Plant Journal, 4, 640-655

Faergeman 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, 37051-37059

Faergeman 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, 1-12

Fulda M, Heinz E,Wolter F P. 1997. Brassica napus cDNAsencoding fatty acyl-CoA synthetase. Plant Molecular Biology, 5, 911-922

Fulda 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, 394-405

Gietz 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, 1,1-4

Groot P H, Scholte H R, Hulsmann W C. 1976. Fattyacid activation: specificity, localization, and function.Advances in Lipid Research, 14, 75-126

Harwood J L. 1996. Recent advances in the biosynthesis ofplant fatty acids. Biochimica et Biophysica Acta, 1301,7-56

Hayashi 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, 2019-2026

Igal 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, 529-534

Jessen 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, 715-726

Karimi M, Inze D, Depicker A. 2002. GATEWAY vectorsfor Agrobacterium-mediated plant transformation.Trends in Plant Science, 5, 193-195

Li 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, 15973-15978

Lu 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, 553-564

Mashek 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, 1958-1961

Ohlrogge J, Browse J. 1995. Lipid biosynthesis. The PlantCell, 7, 957-970

Ohlrogge J B. 1994. Design of new plant products: engineering of fatty acid metabolism. Plant Physiology, 3,821-826

Pongdontri P, Hills M. 2001. Characterization of a novel plant acyl-coA synthetase that is expressed in lipogenictissues of Brassica napus L. Plant Molecular Biology, 6,717-726

Saitou N, Nei M. 1987. The neighbor-joining method: a newmethod for reconstructing phylogenetic trees. MolecularBiology and Evolution, 4, 406-425

Schnurr J, Shockey J, Browse J. 2004. The acyl-CoA synthetase encoded by LACS2 is essential for normal cuticle development in Arabidopsis. The Plant Cell, 3,629-642

Schnurr 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, 1700-1709

Shockey 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, 1710-1722

Slabas A R, Fawcett T. 1992. The biochemistry andmolecular biology of plant lipid biosynthesis. PlantMolecular Biology, 1, 169-191

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 8,1596-1599

Tang 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, 6581-6586

Thakur 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, 252-254

Thelen J J, Ohlrogge J B. 2002. Metabolic engineering of fatty acid biosynthesis in plants. Metabolic Engineering,1, 12-21

Tonon 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, 402-408

Trenkamp 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, 11903-11908

Wang 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, 473-486

Weng H, Molina I, Shockey J, Browse J. 2010. Organ fusioand defective cuticle function in a lacs1 lacs2 double mutant of Arabidopsis. Planta, 5, 1089-1100

Wood 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, 914-924

Zhao 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, 1048-1058

Zheng 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, 1467-1481

Zubenko 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
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