Abstract
In Arabidopsis thaliana, acyl-CoA-binding proteins (ACBPs) are encoded by six genes, and they display varying affinities for acyl-CoA esters. Recombinant ACBP4 and ACBP5 have been shown to bind oleoyl-CoA esters in vitro. In this study, the subcellular localizations of ACBP4 and ACBP5 were determined by biochemical fractionation followed by western blot analyses using anti-ACBP4 and anti-ACBP5 antibodies and immuno-electron microscopy. Confocal microscopy of autofluorescence-tagged ACBP4 and ACBP5, expressed transiently in onion epidermal cells and in transgenic Arabidopsis, confirmed their expression in the cytosol. Taken together, ACBP4 and ACBP5 are available in the cytosol to bind and transfer cytosolic oleoyl-CoA esters. Lipid profile analysis further revealed that an acbp4 knockout mutant showed decreases in membrane lipids (digalactosyldiacylglycerol, monogalactosyldiacylglycerol, phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol) while acbp4-complemented lines attained levels similar to wild type, suggesting that ACBP4 plays a role in the biosynthesis of membrane lipids including galactolipids and phospholipids.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- ACBP:
-
Acyl-coenzyme-A-binding protein
- DGDG:
-
Digalactosyldiacylglycerol
- MGDG:
-
Monogalactosyldiacylglycerol
- PA:
-
Phosphatidic acid
- PC:
-
Phosphatidylcholine
- PE:
-
Phosphatidylethanolamine
- PI:
-
Phosphatidylinositol
References
Abel S, Theologis A (1994) Transient transformation of Arabidopsis leaf protoplasts: a versatile experimental system to study gene expression. Plant J 5:421–427. doi:10.1111/j.1365-313X.1994.00421.x
Bates PD, Ohlrogge JB, Pollard M (2007) Incorporation of newly-synthesized fatty acids into cytosolic glycerolipids in pea leaves occurs via acyl editing. J Biol Chem 282:31206–31216. doi:10.1074/jbc.M705447200
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. doi:10.1016/0003-2697(76)90527-3
Branen JK, Shintani DK, Engeseth NJ (2003) Expression of antisense acyl carrier protein-4 reduces lipid content in Arabidopsis leaf tissue. Plant Physiol 132:748–756. doi:10.1104/pp.102.018622
Browse J, Warwick N, Somerville CR, Slack CR (1986) Fluxes through the prokaryotic and eukaryotic pathways of lipid synthesis in the ‘16:3’ plant Arabidopsis thaliana. Biochem J 235:25–31
Burton M, Rose TM, Faergeman NJ, Knudsen J (2005) Evolution of the acyl-CoA binding protein (ACBP). Biochem J 392:299–307. doi:10.1042/BJ20050829
Chye ML (1998) Arabidopsis cDNA encoding a membrane-associated protein with an acyl-CoA binding domain. Plant Mol Biol 38:827–838. doi:10.1023/A:1006052108468
Chye ML, Huang BQ, Zee SY (1999) Isolation of a gene encoding Arabidopsis membrane-associated acyl-CoA binding protein and immunolocalization of its gene product. Plant J 18:205–214. doi:10.1046/j.1365-313X.1999.00443.x
Chye ML, Li HY, Yung MH (2000) Single amino acid substitutions at the acyl-CoA-binding domain interrupt 14[C]palmitoyl-CoA binding of ACBP2, an Arabidopsis acyl-CoA-binding protein with ankyrin repeats. Plant Mol Biol 44:711–721. doi:10.1023/A:1026524108095
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743. doi:10.1046/j.1365-313x.1998.00343.x
Dörmann P, Balbo I, Benning C (1999) Arabidopsis galactolipid biosynthesis and lipid trafficking mediated by DGD1. Science 284:2181–2184. doi:10.1126/science.284.5423.2181
Engeseth NJ, Pacovsky RS, Newman T, Ohlrogge JB (1996) Characterization of an acyl-CoA-binding protein from Arabidopsis thaliana. Arch Biochem Biophys 331:55–62. doi:10.1006/abbi.1996.0282
Goodin MM, Dietzgen RG, Schichnes D, Ruzin S, Jackson AO (2002) pGD vectors: versatile tools for the expression of green and red fluorescent protein fusions in agroinfiltrated plant leaves. Plant J 31:375–383. doi:10.1046/j.1365-313X.2002.01360.x
Kobayashi K, Kondo M, Fukuda H, Nishimura M, Ohta H (2007) Galactolipid synthesis in chloroplast inner envelope is essential for proper thylakoid biogenesis, photosynthesis, and embryogenesis. Proc Natl Acad Sci USA 104:17216–17221. doi:10.1073/pnas.0704680104
Leung KC, Li HY, Mishra G, Chye ML (2004) ACBP4 and ACBP5, novel Arabidopsis acyl-CoA-binding proteins with kelch motifs that bind oleoyl-CoA. Plant Mol Biol 55:297–309. doi:10.1007/s11103-004-0642-z
Leung KC, Li HY, Xiao S, Tse MH, Chye ML (2006) Arabidopsis ACBP3 is an extracellularly targeted acyl-CoA-binding protein. Planta 223:871–881. doi:10.1007/s00425-005-0139-2
Li HY, Chye ML (2003) Membrane localization of Arabidopsis acyl-CoA binding protein ACBP2. Plant Mol Biol 51:483–492. doi:10.1023/A:1022330304402
Li HY, Chye ML (2004) Arabidopsis acyl-CoA binding protein ACBP2 interacts with an ethylene-responsive element binding protein AtEBP via its ankyrin repeats. Plant Mol Biol 54:233–243. doi:10.1023/B:PLAN.0000028790.75090.ab
Mou Z, He Y, Dai Y, Liu X, Li J (2000) Deficiency in fatty acid synthase leads to premature cell death and dramatic alterations in plant morphology. Plant Cell 12:405–418
Mylne J, Botella JR (1998) Binary vectors for sense and antisense expression of Arabidopsis ESTs. Plant Mol Biol Rep 16:257–262. doi:10.1023/A:1007520410190
Nakai K, Horton P (1999) PSORT: a program for detecting sorting signals in proteins and predicting their subcellular localization. Trends Biochem Sci 24:34–36. doi:10.1016/S0968-0004(98)01336-X
Ohlrogge JB, Browse J (1995) Lipid biosynthesis. Plant Cell 7:957–970
Shi DQ, Liu J, Xiang YH, Ye D, Sundaresan V, Yang WC (2005) SLOW WALKER1, essential for gametogenesis in Arabidopsis, encodes a WD40 protein involved in 18S ribosomal RNA biogenesis. Plant Cell 17:2340–2354. doi:10.1105/tpc.105.033563
Smith JA, Krauss MR, Borkird C, Sung ZR (1988) A nuclear-protein associated with cell divisions in plants. Planta 174:462–472. doi:10.1007/BF00634474
Welti R, Li W, Li M, Sang Y, Biesiada H, Zhou HE, Rajashekar CB, Williams TD, Wang X (2002) Profiling membrane lipids in plant stress responses. Role of phospholipase D alpha in freezing-induced lipid changes in Arabidopsis. J Biol Chem 277:31994–32002. doi:10.1074/jbc.M205375200
Xiao S, Gao W, Chen QF, Ramalingam S, Chye ML (2008) Overexpression of membrane-associated acyl-CoA-binding protein ACBP1 enhances lead tolerance in Arabidopsis. Plant J 54:141–151. doi:10.1111/j.1365-313X.2008.03402.x
Xu ZF, Teng WL, Chye ML (2004) Inhibition of endogenous trypsin- and chymotrypsin-like activities in transgenic lettuce expressing heterogeneous proteinase inhibitor SaPIN2a. Planta 218:623–629. doi:10.1007/s00425-003-1138-9
Zhang Y, Yang C, Li Y, Zheng N, Chen H, Zhao Q, Gao T, Guo H, Xie Q (2007) SDIR1 Is a RING finger E3 ligase that positively regulates stress-responsive abscisic acid signaling in Arabidopsis. Plant Cell 19:1912–1929. doi:10.1105/tpc.106.048488
Acknowledgements
We thank Drs. M.M. Goodin (University of California, Berkeley) and W.C. Yang (Institute of Genetics and Developmental Biology, Beijing) for provision of vectors pGDG and pBI-eGFP, respectively, J. Tam for technical assistance in ultra-thin sectioning, Mary Roth (Kansas Lipidomics Research Center) for lipid profiling, Dr. W.K. Yip for provision of the Biolistic PDS-1000/He system, and Dr. S.F. Sin for generation of construct pAT324 used in complementation of the acbp4 mutant. This work was supported by the Research Grants Council of the Hong Kong Special Administrative Region, China (project HKU7504/05 M), the University of Hong Kong (grant no. 10208034) and the Croucher Foundation (MLC, HL). SX, JZ and SC were supported by postgraduate studentships from the University of Hong Kong. The Kansas Lipidomics Research Center where lipid analysis was carried out was supported by National Science Foundation (EPS 0236913, MCB 0455318, DBI 0521587), Kansas Technology Enterprise Corporation, Kansas IDeA Network of Biomedical Research Excellence (INBRE) of National Institute of Health (P20RR16475), and Kansas State University.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Xiao, S., Li, HY., Zhang, JP. et al. Arabidopsis acyl-CoA-binding proteins ACBP4 and ACBP5 are subcellularly localized to the cytosol and ACBP4 depletion affects membrane lipid composition. Plant Mol Biol 68, 571–583 (2008). https://doi.org/10.1007/s11103-008-9392-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11103-008-9392-7