title: Transcription factor Reb1p regulates DGK1-encoded diacylglycerol kinase and lipid metabolism in Saccharomyces cerevisiae.
abstract: In the yeast Saccharomyces cerevisiae, the DGK1-encoded diacylglycerol kinase catalyzes the CTP-dependent phosphorylation of diacylglycerol to form phosphatidate. This enzyme, in conjunction with PAH1-encoded phosphatidate phosphatase, controls the levels of phosphatidate and diacylglycerol for phospholipid synthesis, membrane growth, and lipid droplet formation. In this work, we showed that a functional level of diacylglycerol kinase is regulated by the Reb1p transcription factor. In the electrophoretic mobility shift assay, purified recombinant Reb1p was shown to specifically bind its consensus recognition sequence (CGGGTAA, -166 to -160) in the DGK1 promoter. Analysis of cells expressing the PDGK1-lacZ reporter gene showed that mutations (GT→TG) in the Reb1p-binding sequence caused an 8.6-fold reduction in β-galactosidase activity. The expression of DGK1(reb1), a DGK1 allele containing the Reb1p-binding site mutation, was greatly lower than that of the wild type allele, as indicated by analyses of DGK1 mRNA, Dgk1p, and diacylglycerol kinase activity. In the presence of cerulenin, an inhibitor of de novo fatty acid synthesis, the dgk1Δ mutant expressing DGK1(reb1) exhibited a significant defect in growth as well as in the synthesis of phospholipids from triacylglycerol mobilization. Unlike DGK1, the DGK1(reb1) expressed in the dgk1Δ pah1Δ mutant did not result in the nuclear/endoplasmic reticulum membrane expansion, which occurs in cells lacking phosphatidate phosphatase activity. Taken together, these results indicate that the Reb1p-mediated regulation of diacylglycerol kinase plays a major role in its in vivo functions in lipid metabolism.
authors: From the Department of Food Science, Rutgers Center for Lipid Research, and New Jersey Institute for Food, Nutrition, and Health, Rutgers University, New Brunswick, New Jersey 08901 and.
authors: Yixuan
authors: Y
authors: Qiu
authors: Stylianos
authors: S
authors: Fakas
authors: Gil-Soo
authors: GS
authors: Han
authors: Antonio Daniel
authors: AD
authors: Barbosa
authors: Symeon
authors: S
authors: Siniossoglou
authors: George M
authors: GM
authors: Carman
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005227:Fatty Acids / Q000096:biosynthesis; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D050356:Lipid Metabolism / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010743:Phospholipids / Q000096:biosynthesis; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014280:Triglycerides / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: C434524:DGK1 protein, S cerevisiae; D004268:DNA-Binding Proteins; D005227:Fatty Acids; D010743:Phospholipids; C065681:REB1 protein, S cerevisiae; D012333:RNA, Messenger; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D014280:Triglycerides; D001616:beta-Galactosidase
keywords: Diacylglycerol; Diacylglycerol Kinase; Glycerolipid; Phosphatidate; Phospholipid; Phospholipid Metabolism; Phospholipid Turnover
references: Biochim Biophys Acta. 2007 Jul;1771(7):793-806
references: 17512245
references: Biochim Biophys Acta. 2009 Sep;1791(9):942-8
references: 19264149
references: Methods Enzymol. 1983;96:192-205
references: 6228704
references: J Biol Chem. 2006 Apr 7;281(14):9210-8
references: 16467296
references: Methods Enzymol. 1991;197:548-53
references: 2051943
references: J Biol Chem. 2000 Jun 30;275(26):19475-81
references: 10777505
references: Gene. 1995 Aug 8;161(1):75-9
references: 7642141
references: Prog Lipid Res. 2006 May;45(3):250-78
references: 16574237
references: Curr Drug Targets. 2008 Aug;9(8):626-40
references: 18691010
references: J Biol Chem. 1999 Apr 23;274(17):11447-50
references: 10206945
references: J Biol Chem. 1987 Mar 25;262(9):4231-40
references: 3031066
references: J Biol Chem. 1986 May 25;261(15):6993-7000
references: 3009483
references: EMBO J. 1992 Dec;11(13):5051-61
references: 1464327
references: J Biol Chem. 2009 Oct 2;284(40):27609-19
references: 19656950
references: Genes Dev. 2009 Aug 15;23(16):1929-43
references: 19684113
references: EMBO J. 2005 Jun 1;24(11):1931-41
references: 15889145
references: J Biol Chem. 2000 Dec 29;275(52):40887-96
references: 11016943
references: J Biol Chem. 2011 Aug 19;286(33):29074-29085
references: 21708942
references: J Biol Chem. 2003 Jun 27;278(26):23317-23
references: 12682047
references: Mol Gen Genet. 1986 Jun;203(3):479-86
references: 3528750
references: Cancer Res. 2007 Jan 1;67(1):1-4
references: 17210675
references: Mol Gen Genet. 1996 Feb 25;250(3):348-56
references: 8602150
references: J Biol Chem. 2011 Jan 14;286(2):1464-74
references: 21071438
references: J Biol Chem. 1993 May 25;268(15):10946-52
references: 8098706
references: Can J Biochem Physiol. 1959 Aug;37(8):911-7
references: 13671378
references: J Biol Chem. 2008 Apr 11;283(15):10079-88
references: 18276583
references: Biochim Biophys Acta. 2013 Mar;1831(3):575-81
references: 23026159
references: Nature. 1970 Aug 15;227(5259):680-5
references: 5432063
references: J Biol Chem. 2010 May 21;285(21):15663-7
references: 20231294
references: Proc Natl Acad Sci U S A. 1992 May 15;89(10):4534-8
references: 1350093
references: Yeast. 1994 Jun;10(6):771-87
references: 7975895
references: Trends Plant Sci. 2005 Aug;10(8):368-75
references: 16023886
references: J Biol Chem. 2006 Jan 6;281(1):491-500
references: 16267052
references: J Proteome Res. 2007 Mar;6(3):1190-7
references: 17330950
references: Prog Lipid Res. 2008 May;47(3):157-71
references: 18258205
references: Trends Biochem Sci. 2007 Jan;32(1):27-36
references: 17157506
references: Proc Natl Acad Sci U S A. 2010 Oct 12;107(41):17539-44
references: 20876142
references: Nature. 1997 May 1;387(6628):101-5
references: 9139830
references: J Biol Chem. 1990 Jan 25;265(3):1235-8
references: 2404013
references: FEBS J. 2008 Nov;275(22):5552-63
references: 18959743
references: Anal Biochem. 1976 May 7;72:248-54
references: 942051
references: Mol Microbiol. 1995 Nov;18(4):605-14
references: 8817483
references: J Biol Chem. 2008 Jul 18;283(29):20443-53
references: 18458076
references: Mol Gen Genet. 1997 Nov;256(6):682-9
references: 9435793
references: Methods Enzymol. 1991;194:398-405
references: 1706459
references: J Biol Chem. 1965 Jun;240:2468-77
references: 14304855
references: Mol Cell Biol. 1990 May;10(5):2269-84
references: 2183028
references: Nucleic Acids Res. 1996 Dec 1;24(23):4599-607
references: 8967907
references: BMC Genomics. 2010 Dec 02;11:685
references: 21126336
references: J Biol Chem. 2008 Jul 18;283(29):20433-42
references: 18458075
references: Mol Cell Biol. 1998 Jul;18(7):4368-76
references: 9632820
references: Biochim Biophys Acta. 2002 May 23;1582(1-3):45-51
references: 12069809
references: J Cell Biol. 2011 Mar 21;192(6):1043-55
references: 21422231
references: Mol Cell Biol. 1990 Oct;10(10):5226-34
references: 2204808
references: Methods Enzymol. 1981;72:520-32
references: 7031426
references: Mol Cell Biol. 1990 Apr;10(4):1743-53
references: 2181283
references: Biochim Biophys Acta. 1999 Jul 30;1439(2):299-316
references: 10425403
references: Biochim Biophys Acta. 2013 Mar;1831(3):514-22
references: 22910056
references: Crit Rev Biochem Mol Biol. 2011 Oct;46(5):353-64
references: 21539478
references: J Biol Chem. 2005 Nov 11;280(45):37301-9
references: 16135509
references: Mol Biosyst. 2009 Nov;5(11):1337-46
references: 19823750
references: J Cell Biochem. 2004 Aug 1;92(5):900-12
references: 15258914
references: J Biol Chem. 2012 Jan 13;287(3):2221-36
references: 22121197
references: J Biol Chem. 2009 Nov 6;284(45):30994-1005
references: 19690167
references: Chem Rev. 2011 Oct 12;111(10):6186-208
references: 21800853
references: Mol Microbiol. 1994 Jun;12(6):931-40
references: 7934900
references: Nucleic Acids Res. 1990 May 25;18(10):3091-2
references: 2190191
references: J Bacteriol. 1983 Jan;153(1):163-8
references: 6336730
references: Eur J Biochem. 1994 Oct 1;225(1):213-22
references: 7925441
references: J Biol Chem. 2005 Nov 18;280(46):38328-36
references: 16179339
references: Cancer Discov. 2013 Jul;3(7):782-97
references: 23558954
references: J Biol Chem. 1994 Apr 1;269(13):9790-7
references: 8144571
references: Front Immunol. 2013 Jul 04;4:178
references: 23847619
references: Chem Phys Lipids. 1999 Apr;98(1-2):109-17
references: 10358933
references: Annu Rev Biochem. 2011;80:859-83
references: 21275641
references: Genetics. 1989 May;122(1):19-27
references: 2659436
references: J Biol Chem. 1990 Dec 5;265(34):20778-83
references: 2249986
references: Front Biosci. 2008 Jan 01;13:590-7
references: 17981572
references: J Biol Chem. 2007 Dec 28;282(52):37293-7
references: 17981800
references: J Biol Chem. 2006 Nov 10;281(45):34537-48
references: 16968695
references: J Biol Chem. 2007 Dec 21;282(51):37026-35
references: 17971454
references: Anal Biochem. 1981 Apr;112(2):195-203
references: 6266278
references: Mol Cell Biol. 1992 Dec;12(12):5516-26
references: 1448083
references: Proteomics. 2009 Oct;9(20):4642-52
references: 19795423
references: Mol Gen Genet. 1994 Jul 8;244(1):90-6
references: 8041367
references: Mol Cell. 2009 Jan 16;33(1):53-63
references: 19150427
references: Genetics. 2012 Feb;190(2):317-49
references: 22345606
references: Biochem J. 2008 Jan 1;409(1):1-18
references: 18062770
references: Can J Physiol Pharmacol. 2006 Jan;84(1):29-38
references: 16845888
references: J Biol Chem. 1983 Oct 10;258(19):11648-53
references: 6311818
references: Leuk Res. 2001 Oct;25(10):891-900
references: 11532523
references: Prog Lipid Res. 2012 Jan;51(1):1-10
references: 22067957
references: Biochim Biophys Acta. 2007 Mar;1771(3):255-70
references: 16950653
references: Biochim Biophys Acta. 1995 Mar 22;1234(2):214-20
references: 7696296
references: Prog Lipid Res. 1999 Sep-Nov;38(5-6):361-99
references: 10793889
references: Appl Environ Microbiol. 1990 Jun;56(6):1875-81
references: 16348228
references: Mol Cell Biol. 1993 Jan;13(1):649-58
references: 8417359
references: Genes Dev. 1990 Apr;4(4):503-14
references: 2361590