Abstract
A putative yeast sn-2 acyltransferase gene (SLC1-1), reportedly a variant acyltransferase that suppresses a genetic defect in sphingolipid long-chain base biosynthesis, has been expressed in a yeast SLC deletion strain. The SLC1-1 gene product was shown in vitro to encode an sn-2 acyltransferase capable of acylating sn-1 oleoyl-lysophosphatidic acid, using a range of acyl-CoA thioesters, including 18:1-, 22:1-, and 24:0-CoAs. The SLC1-1 gene was introduced into Arabidopsis and a high erucic acid-containing Brassica napus cv Hero under the control of a constitutive (tandem cauliflower mosaic virus 35S) promoter. The resulting transgenic plants showed substantial increases of 8 to 48% in seed oil content (expressed on the basis of seed dry weight) and increases in both overall proportions and amounts of very-long-chain fatty acids in seed triacylglycerols (TAGs). Furthermore, the proportion of very-long-chain fatty acids found at the sn-2 position of TAGs was increased, and homogenates prepared from developing seeds of transformed plants exhibited elevated lysophosphatidic acid acyltransferase (EC 2.3.1.51) activity. Thus, the yeast sn-2 acyltransferase has been shown to encode a protein that can exhibit lysophosphatidic acid acyltransferase activity and that can be used to change total fatty acid content and composition as well as to alter the stereospecific acyl distribution of fatty acids in seed TAGs.
Full Text
The Full Text of this article is available as a PDF (1.8 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Brown A. P., Brough C. L., Kroon J. T., Slabas A. R. Identification of a cDNA that encodes a 1-acyl-sn-glycerol-3-phosphate acyltransferase from Limnanthes douglasii. Plant Mol Biol. 1995 Oct;29(2):267–278. doi: 10.1007/BF00043651. [DOI] [PubMed] [Google Scholar]
- Carman G. M., Zeimetz G. M. Regulation of phospholipid biosynthesis in the yeast Saccharomyces cerevisiae. J Biol Chem. 1996 Jun 7;271(23):13293–13296. doi: 10.1074/jbc.271.23.13293. [DOI] [PubMed] [Google Scholar]
- Coleman J. Characterization of Escherichia coli cells deficient in 1-acyl-sn-glycerol-3- phosphate acyltransferase activity. J Biol Chem. 1990 Oct 5;265(28):17215–17221. [PubMed] [Google Scholar]
- Datla R. S., Hammerlindl J. K., Panchuk B., Pelcher L. E., Keller W. Modified binary plant transformation vectors with the wild-type gene encoding NPTII. Gene. 1992 Dec 15;122(2):383–384. doi: 10.1016/0378-1119(92)90232-e. [DOI] [PubMed] [Google Scholar]
- De Block M., De Brouwer D., Tenning P. Transformation of Brassica napus and Brassica oleracea Using Agrobacterium tumefaciens and the Expression of the bar and neo Genes in the Transgenic Plants. Plant Physiol. 1989 Oct;91(2):694–701. doi: 10.1104/pp.91.2.694. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elble R. A simple and efficient procedure for transformation of yeasts. Biotechniques. 1992 Jul;13(1):18–20. [PubMed] [Google Scholar]
- Erickson R. P., Hudgins L., Stone J. F., Schmidt S., Wilke C., Glover T. W. A "balanced" Y;16 translocation associated with Turner-like neonatal lymphedema suggests the location of a potential anti-Turner gene on the Y chromosome. Cytogenet Cell Genet. 1995;71(2):163–167. doi: 10.1159/000134099. [DOI] [PubMed] [Google Scholar]
- Gavey K. L., Trujillo D. L., Scallen T. J. Evidence for phosphorylation/dephosphorylation of rat liver acyl-CoA:cholesterol acyltransferase. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2171–2174. doi: 10.1073/pnas.80.8.2171. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Katavic V., Haughn G. W., Reed D., Martin M., Kunst L. In planta transformation of Arabidopsis thaliana. Mol Gen Genet. 1994 Nov 1;245(3):363–370. doi: 10.1007/BF00290117. [DOI] [PubMed] [Google Scholar]
- Katavic V., Reed D. W., Taylor D. C., Giblin E. M., Barton D. L., Zou J., Mackenzie S. L., Covello P. S., Kunst L. Alteration of seed fatty acid composition by an ethyl methanesulfonate-induced mutation in Arabidopsis thaliana affecting diacylglycerol acyltransferase activity. Plant Physiol. 1995 May;108(1):399–409. doi: 10.1104/pp.108.1.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knutzon D. S., Lardizabal K. D., Nelsen J. S., Bleibaum J. L., Davies H. M., Metz J. G. Cloning of a coconut endosperm cDNA encoding a 1-acyl-sn-glycerol-3-phosphate acyltransferase that accepts medium-chain-length substrates. Plant Physiol. 1995 Nov;109(3):999–1006. doi: 10.1104/pp.109.3.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knutzon D. S., Thompson G. A., Radke S. E., Johnson W. B., Knauf V. C., Kridl J. C. Modification of Brassica seed oil by antisense expression of a stearoyl-acyl carrier protein desaturase gene. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2624–2628. doi: 10.1073/pnas.89.7.2624. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lassner M. W., Lardizabal K., Metz J. G. A jojoba beta-Ketoacyl-CoA synthase cDNA complements the canola fatty acid elongation mutation in transgenic plants. Plant Cell. 1996 Feb;8(2):281–292. doi: 10.1105/tpc.8.2.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lassner M. W., Levering C. K., Davies H. M., Knutzon D. S. Lysophosphatidic acid acyltransferase from meadowfoam mediates insertion of erucic acid at the sn-2 position of triacylglycerol in transgenic rapeseed oil. Plant Physiol. 1995 Dec;109(4):1389–1394. doi: 10.1104/pp.109.4.1389. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lester R. L., Wells G. B., Oxford G., Dickson R. C. Mutant strains of Saccharomyces cerevisiae lacking sphingolipids synthesize novel inositol glycerophospholipids that mimic sphingolipid structures. J Biol Chem. 1993 Jan 15;268(2):845–856. [PubMed] [Google Scholar]
- Lindstrom J. T., Vodkin L. O. A soybean cell wall protein is affected by seed color genotype. Plant Cell. 1991 Jun;3(6):561–571. doi: 10.1105/tpc.3.6.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mol J. N., van der Krol A. R., van Tunen A. J., van Blokland R., de Lange P., Stuitje A. R. Regulation of plant gene expression by antisense RNA. FEBS Lett. 1990 Aug 1;268(2):427–430. doi: 10.1016/0014-5793(90)81298-3. [DOI] [PubMed] [Google Scholar]
- Nagiec M. M., Wells G. B., Lester R. L., Dickson R. C. A suppressor gene that enables Saccharomyces cerevisiae to grow without making sphingolipids encodes a protein that resembles an Escherichia coli fatty acyltransferase. J Biol Chem. 1993 Oct 15;268(29):22156–22163. [PubMed] [Google Scholar]
- Stark D. M., Timmerman K. P., Barry G. F., Preiss J., Kishore G. M. Regulation of the Amount of Starch in Plant Tissues by ADP Glucose Pyrophosphorylase. Science. 1992 Oct 9;258(5080):287–292. doi: 10.1126/science.258.5080.287. [DOI] [PubMed] [Google Scholar]
- Taylor D. C., Barton D. L., Giblin E. M., MacKenzie S. L., Van Den Berg CGJ., McVetty PBE. Microsomal Lyso-Phosphatidic Acid Acyltransferase from a Brassica oleracea Cultivar Incorporates Erucic Acid into the sn-2 Position of Seed Triacylglycerols. Plant Physiol. 1995 Oct;109(2):409–420. doi: 10.1104/pp.109.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor D. C., Barton D. L., Rioux K. P., Mackenzie S. L., Reed D. W., Underhill E. W., Pomeroy M. K., Weber N. Biosynthesis of Acyl Lipids Containing Very-Long Chain Fatty Acids in Microspore-Derived and Zygotic Embryos of Brassica napus L. cv Reston. Plant Physiol. 1992 Aug;99(4):1609–1618. doi: 10.1104/pp.99.4.1609. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor D. C., Weber N., Hogge L. R., Underhill E. W. A simple enzymatic method for the preparation of radiolabeled erucoyl-CoA and other long-chain fatty acyl-CoAs and their characterization by mass spectrometry. Anal Biochem. 1990 Feb 1;184(2):311–316. doi: 10.1016/0003-2697(90)90686-4. [DOI] [PubMed] [Google Scholar]
- Voelker T. A., Worrell A. C., Anderson L., Bleibaum J., Fan C., Hawkins D. J., Radke S. E., Davies H. M. Fatty acid biosynthesis redirected to medium chains in transgenic oilseed plants. Science. 1992 Jul 3;257(5066):72–74. doi: 10.1126/science.1621095. [DOI] [PubMed] [Google Scholar]