HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ostrander, D. B. Articles by Carman, G. M. Search for Related Content PubMed PubMed Citation Articles by Ostrander, D. B. Articles by Carman, G. M. Social Bookmarking                      What's this?

J Biol Chem, Vol. 273, Issue 30, 18992-19001, July 24, 1998

Effect of CTP Synthetase Regulation by CTP on Phospholipid Synthesis in Saccharomyces cerevisiae

Darin B. Ostrander, Daniel J. O'Brien, Jessica A. Gorman^§, and George M. Carman

From the  Department of Food Science, Cook College, New Jersey Agricultural Experiment Station, Rutgers University, New Brunswick, New Jersey 08901 and the ^§ Department of Microbial Molecular Biology, Pharmaceutical Research Institute, Bristol-Myers Squibb, Princeton, New Jersey 08543

CTP synthetase (EC 6.3.4.2, UTP:ammonia ligase (ADP-forming)) activity in Saccharomyces cerevisiae is allosterically regulated by CTP product inhibition. Amino acid residue Glu¹⁶¹ in the URA7-encoded and URA8-encoded CTP synthetases was identified as being involved in the regulation of these enzymes by CTP product inhibition. The specific activities of the URA7-encoded and URA8-encoded enzymes with a Glu¹⁶¹  Lys (E161K) mutation were 2-fold greater when compared with the wild-type enzymes. The E161K mutant URA7-encoded and URA8-encoded CTP synthetases were less sensitive to CTP product inhibition with inhibitor constants for CTP of 8.4- and 5-fold greater, respectively, than those of their wild-type counterparts. Cells expressing the E161K mutant enzymes on a multicopy plasmid exhibited an increase in resistance to the pyrimidine poison and cancer therapeutic drug cyclopentenylcytosine and accumulated elevated (6-15-fold) levels of CTP when compared with cells expressing the wild-type enzymes. Cells expressing the E161K mutation in the URA7-encoded CTP synthetase exhibited an increase (1.5-fold) in the utilization of the Kennedy pathway for phosphatidylcholine synthesis when compared with control cells. Cells bearing the mutation also exhibited an increase in the synthesis of phosphatidylcholine (1.5-fold), phosphatidylethanolamine (1.3-fold), and phosphatidate (2-fold) and a decrease in the synthesis of phosphatidylserine (1.7-fold). These alterations were accompanied by an inositol excretion phenotype due to the misregulation of the INO1 gene. Moreover, cells bearing the E161K mutation exhibited an increase (1.6-fold) in the ratio of total neutral lipids to phospholipids, an increase in triacylglycerol (1.4-fold), free fatty acids (1.7-fold), and ergosterol ester (1.8-fold), and a decrease in diacylglycerol (1.3-fold) when compared with control cells. These data indicated that the regulation of CTP synthetase activity by CTP plays an important role in the regulation of phospholipid synthesis.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				G. M. Carman and G.-S. Han Regulation of phospholipid synthesis in yeast J. Lipid Res., April 1, 2009; 50(Supplement): S69 - S73. [Abstract] [Full Text] [PDF]

				G. M. Carman and G.-S. Han Phosphatidic Acid Phosphatase, a Key Enzyme in the Regulation of Lipid Synthesis J. Biol. Chem., January 30, 2009; 284(5): 2593 - 2597. [Full Text] [PDF]

				G.-S. Han, L. O'Hara, S. Siniossoglou, and G. M. Carman Characterization of the Yeast DGK1-encoded CTP-dependent Diacylglycerol Kinase J. Biol. Chem., July 18, 2008; 283(29): 20443 - 20453. [Abstract] [Full Text] [PDF]

				G.-S. Han, L. O'Hara, G. M. Carman, and S. Siniossoglou An Unconventional Diacylglycerol Kinase That Regulates Phospholipid Synthesis and Nuclear Membrane Growth J. Biol. Chem., July 18, 2008; 283(29): 20433 - 20442. [Abstract] [Full Text] [PDF]

				F. A. Lunn, J. E. MacDonnell, and S. L. Bearne Structural Requirements for the Activation of Escherichia coli CTP Synthase by the Allosteric Effector GTP Are Stringent, but Requirements for Inhibition Are Lax J. Biol. Chem., January 25, 2008; 283(4): 2010 - 2020. [Abstract] [Full Text] [PDF]

				G. M. Carman and S. A. Henry Phosphatidic Acid Plays a Central Role in the Transcriptional Regulation of Glycerophospholipid Synthesis in Saccharomyces cerevisiae J. Biol. Chem., December 28, 2007; 282(52): 37293 - 37297. [Full Text] [PDF]

				G.-S. Han, S. Siniossoglou, and G. M. Carman The Cellular Functions of the Yeast Lipin Homolog Pah1p Are Dependent on Its Phosphatidate Phosphatase Activity J. Biol. Chem., December 21, 2007; 282(51): 37026 - 37035. [Abstract] [Full Text] [PDF]

				H.-S. Choi and G. M. Carman Respiratory Deficiency Mediates the Regulation of CHO1-encoded Phosphatidylserine Synthase by mRNA Stability in Saccharomyces cerevisiae J. Biol. Chem., October 26, 2007; 282(43): 31217 - 31227. [Abstract] [Full Text] [PDF]

				Y. Nakamura, M. Tsuchiya, and H. Ohta Plastidic Phosphatidic Acid Phosphatases Identified in a Distinct Subfamily of Lipid Phosphate Phosphatases with Prokaryotic Origin J. Biol. Chem., September 28, 2007; 282(39): 29013 - 29021. [Abstract] [Full Text] [PDF]

				Y.-F. Chang, S. S. Martin, E. P. Baldwin, and G. M. Carman Phosphorylation of Human CTP Synthetase 1 by Protein Kinase C: IDENTIFICATION OF Ser462 AND Thr455 AS MAJOR SITES OF PHOSPHORYLATION J. Biol. Chem., June 15, 2007; 282(24): 17613 - 17622. [Abstract] [Full Text] [PDF]

				M.-G. Choi and G. M. Carman Phosphorylation of Human CTP Synthetase 1 by Protein Kinase A: IDENTIFICATION OF Thr455 AS A MAJOR SITE OF PHOSPHORYLATION J. Biol. Chem., February 23, 2007; 282(8): 5367 - 5377. [Abstract] [Full Text] [PDF]

				J. Singh, D. Kumar, N. Ramakrishnan, V. Singhal, J. Jervis, J. F. Garst, S. M. Slaughter, A. M. DeSantis, M. Potts, and R. F. Helm Transcriptional Response of Saccharomyces cerevisiae to Desiccation and Rehydration Appl. Envir. Microbiol., December 1, 2005; 71(12): 8752 - 8763. [Abstract] [Full Text] [PDF]

				G.-S. Han, A. Sreenivas, M.-G. Choi, Y.-F. Chang, S. S. Martin, E. P. Baldwin, and G. M. Carman Expression of Human CTP Synthetase in Saccharomyces cerevisiae Reveals Phosphorylation by Protein Kinase A J. Biol. Chem., November 18, 2005; 280(46): 38328 - 38336. [Abstract] [Full Text] [PDF]

				M.-G. Choi, V. Kurnov, M. C. Kersting, A. Sreenivas, and G. M. Carman Phosphorylation of the Yeast Choline Kinase by Protein Kinase C: IDENTIFICATION OF Ser25 AND Ser30 AS MAJOR SITES OF PHOSPHORYLATION J. Biol. Chem., July 15, 2005; 280(28): 26105 - 26112. [Abstract] [Full Text] [PDF]

				W. M. Iwanyshyn, G.-S. Han, and G. M. Carman Regulation of Phospholipid Synthesis in Saccharomyces cerevisiae by Zinc J. Biol. Chem., May 21, 2004; 279(21): 21976 - 21983. [Abstract] [Full Text] [PDF]

				H.-S. Choi, A. Sreenivas, G.-S. Han, and G. M. Carman Regulation of Phospholipid Synthesis in the Yeast cki1{Delta} eki1{Delta} Mutant Defective in the Kennedy Pathway: THE CHO1-ENCODED PHOSPHATIDYLSERINE SYNTHASE IS REGULATED BY mRNA STABILITY J. Biol. Chem., March 26, 2004; 279(13): 12081 - 12087. [Abstract] [Full Text] [PDF]

				M.-G. Choi, T.-S. Park, and G. M. Carman Phosphorylation of Saccharomyces cerevisiae CTP Synthetase at Ser424 by Protein Kinases A and C Regulates Phosphatidylcholine Synthesis by the CDP-choline Pathway J. Biol. Chem., June 20, 2003; 278(26): 23610 - 23616. [Abstract] [Full Text] [PDF]

				T.-S. Park, D. J. O'Brien, and G. M. Carman Phosphorylation of CTP Synthetase on Ser36, Ser330, Ser354, and Ser454 Regulates the Levels of CTP and Phosphatidylcholine Synthesis in Saccharomyces cerevisiae J. Biol. Chem., May 30, 2003; 278(23): 20785 - 20794. [Abstract] [Full Text] [PDF]

				M. Willemoes Thr-431 and Arg-433 Are Part of a Conserved Sequence Motif of the Glutamine Amidotransferase Domain of CTP Synthases and Are Involved in GTP Activation of the Lactococcus lactis Enzyme J. Biol. Chem., March 7, 2003; 278(11): 9407 - 9411. [Abstract] [Full Text] [PDF]

				Y. Yu, A. Sreenivas, D. B. Ostrander, and G. M. Carman Phosphorylation of Saccharomyces cerevisiae Choline Kinase on Ser30 and Ser85 by Protein Kinase A Regulates Phosphatidylcholine Synthesis by the CDP-choline Pathway J. Biol. Chem., September 13, 2002; 277(38): 34978 - 34986. [Abstract] [Full Text] [PDF]

				K. Kim, K.-H. Kim, M. K. Storey, D. R. Voelker, and G. M. Carman Isolation and Characterization of the Saccharomyces cerevisiae EKI1 Gene Encoding Ethanolamine Kinase J. Biol. Chem., May 21, 1999; 274(21): 14857 - 14866. [Abstract] [Full Text] [PDF]

				D. B. Ostrander and J. A. Gorman The Extracellular Domain of the Saccharomyces cerevisiae Sln1p Membrane Osmolarity Sensor Is Necessary for Kinase Activity J. Bacteriol., April 15, 1999; 181(8): 2527 - 2534. [Abstract] [Full Text]

				K.-H. Kim and G. M. Carman Phosphorylation and Regulation of Choline Kinase from Saccharomyces cerevisiae by Protein Kinase A J. Biol. Chem., April 2, 1999; 274(14): 9531 - 9538. [Abstract] [Full Text] [PDF]

				S. F. O'Handley, C. A. Dunn, and M. J. Bessman Orf135 from Escherichia coli Is a Nudix Hydrolase Specific for CTP, dCTP, and 5-Methyl-dCTP J. Biol. Chem., February 16, 2001; 276(8): 5421 - 5426. [Abstract] [Full Text] [PDF]