Cis and trans regulatory elements required for regulation of the<i>CHO1</i>gene of<i>Saccharomyces cerevisiae</i>

Bailis, Adam M.; Lopes, John M.; Kohlwein, Sepp D.; Henry, Susan A.

doi:10.1093/nar/20.6.1411

Cited by 63 publications

(68 citation statements)

References 37 publications

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“…CHO1 expression is regulated by water-soluble phospholipid precursors (e.g. inositol supplementation) (59,90,94,95) and by growth phase (96,97). The activity of PS synthase is modulated by membrane phospholipids (98 -100) and is inhibited by inositol (101) and by CTP (39).…”

Section: Discussionmentioning

confidence: 99%

“…Plasmids were maintained and amplified in E. coli strain DH5␣. Plasmid pAB709 (P CHO1 -lacZ) contains 0.3 kilobases of the CHO1 promoter fused to the coding sequence of the E. coli lacZ gene (59). Plasmid pHC2 is a derivative of pAB709, in which the core sequence of the UAS INO element (13) in the CHO1 promoter was changed from 5Ј-CTTTCACAT-3Ј to 5Ј-CTTTAAAAA-3Ј.…”

Section: Methodsmentioning

confidence: 99%

“…Mutant-Maximum expression of CHO1 in wild type cells grown in the absence of inositol is dependent on the UAS INO cis-acting element in its promoter (13,59,79). The UAS INO element contains a consensus-binding site (5Ј-CANNTG-3Ј) for a heterodimer complex of the positive transcription factors Ino2p and Ino4p (13,59,79,89).…”

Section: Effect Of the Uas Ino Element On The Regulation Of Cho1 Exprmentioning

confidence: 99%

“…The UAS INO element contains a consensus-binding site (5Ј-CANNTG-3Ј) for a heterodimer complex of the positive transcription factors Ino2p and Ino4p (13,59,79,89). We questioned whether the UAS INO element played a role in the regulation of PS synthase observed in the cki1⌬ eki1⌬ mutant.…”

Section: Effect Of the Uas Ino Element On The Regulation Of Cho1 Exprmentioning

confidence: 99%

“…The expression of CHO1 in wild type S. cerevisiae is repressed by the addition of inositol to the growth medium (59,90). To examine whether the cki1⌬ eki1⌬ mutations affected this regulation, ␤-galactosidase activity was measured in wild type and mutant cells grown in the absence and presence of 50 M inositol.…”

Section: Effect Of the Uas Ino Element On The Regulation Of Cho1 Exprmentioning

confidence: 99%

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Regulation of Phospholipid Synthesis in the Yeast cki1Δ eki1Δ Mutant Defective in the Kennedy Pathway

Choi

Sreenivas

Han

et al. 2004

Journal of Biological Chemistry

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In the yeast Saccharomyces cerevisiae, the most abundant phospholipid phosphatidylcholine is synthesized by the complementary CDP-diacylglycerol and Kennedy pathways. Using a cki1⌬ eki1⌬ mutant defective in choline kinase and ethanolamine kinase, we examined the consequences of a block in the Kennedy pathway on the regulation of phosphatidylcholine synthesis by the CDP-diacylglycerol pathway. The cki1⌬ eki1⌬ mutant exhibited increases in the synthesis of phosphatidylserine, phosphatidylethanolamine, and phosphatidylcholine via the CDP-diacylglycerol pathway. The increase in phospholipid synthesis correlated with increased activity levels of the CDPdiacylglycerol pathway enzymes phosphatidylserine synthase, phosphatidylserine decarboxylase, phosphatidylethanolamine methyltransferase, and phospholipid methyltransferase. However, other enzyme activities, including phosphatidylinositol synthase and phosphatidate phosphatase, were not affected in the cki1⌬ eki1⌬ mutant. For phosphatidylserine synthase, the enzyme catalyzing the committed step in the pathway, activity was regulated by increases in the levels of mRNA and protein. Decay analysis of CHO1 mRNA indicated that a dramatic increase in transcript stability was a major component responsible for the elevated level of phosphatidylserine synthase. These results revealed a novel mechanism that controls phospholipid synthesis in yeast.PC 1 is the most abundant phospholipid in the membranes of eukaryotic cells (1-4). It serves as a structural component of the membrane and as a source of bioactive lipids (e.g. lyso-PC, PA, DAG) (1-5). The importance of PC is underscored by the fact that alterations in its metabolism are linked to apoptosis (6 -9) and oncogenic transformation (10 -12). In the model eukaryote Saccharomyces cerevisiae, PC is synthesized by complementary pathways that are common to those found in mammalian cells 2 ( Fig. 1) (1,4,(13)(14)(15)(16)(17). In the CDP-DAG pathway, PC is synthesized from CDP-DAG via the reactions catalyzed by PS synthase (18 -20), PS decarboxylase (21-23), PE methyltransferase (24, 25), and phospholipid methyltransferase (24,26). In the CDP-choline branch of the Kennedy pathway, PC is synthesized from choline via the reactions catalyzed by choline kinase (27), phosphocholine cytidylyltransferase (28), and choline phosphotransferase (29,30). Analyses of mutations in S. cerevisiae (4, 31, 32), as well as in mammalian cells (33,34) indicate that the physiological role(s) of PC synthesized by the two pathways are different.The contribution of the CDP-DAG and Kennedy pathways for PC synthesis in wild type S. cerevisiae is dependent on the exogenous supply of choline (35). When grown in the presence of choline, yeast primarily synthesizes PC via the Kennedy pathway (35). On the other hand, when cells are grown in the absence of choline, PC is primarily synthesized via the CDP-DAG pathway (35). Yet, even under this growth condition, the Kennedy pathway still contributes to the synthesis of PC (36 -41). The choline required for the Kenne...

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Effect Of the Uas Ino Element On The Regulation Of Cho1 Exprmentioning

confidence: 99%

Section: Effect Of the Uas Ino Element On The Regulation Of Cho1 Exprmentioning

confidence: 99%

Section: Effect Of the Uas Ino Element On The Regulation Of Cho1 Exprmentioning

confidence: 99%

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Regulation of Phospholipid Synthesis in the Yeast cki1Δ eki1Δ Mutant Defective in the Kennedy Pathway

Choi

Sreenivas

Han

et al. 2004

Journal of Biological Chemistry

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Functional characterization of the repeated UASINO element in the promoters of the INO1 and CHO2 genes of yeast

Koipally

Ashburner

Bachhawat

et al. 1996

Yeast

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In yeast, INO1 and CHO2 gene expression is subject to repression in response to inositol and choline supplementation. The response by both genes to inositol is controlled by a single set of regulatory factors and the highly conserved and repeated UASINO element (consensus: 5′ CATGTGAAAT 3′) that is found in multiple copies in both promoters. However, none of the native elements found in the INO1 and CHO2 promoters constitutes an exact match to the consensus element and the functionality of individual elements from these two promoters has not been tested. In this study, the function of individual putative UASINO elements from both promoters was tested by placing promoter fragments into a reporter construct which lacked a UAS element but contained the TATA element and start of transcription from the yeast CYC1 gene fused to the Escherichia coli lacZ gene. In addition, a set of oligonucleotides containing the consensus UASINO element with the first position systematically modified was also tested for UASINO function. These studies indicated that elements that contain a C or an A as the first base at the 5′ end are functional to varying degrees. The majority of potential UASINO elements from the INO1 promoter were found to be inactive, whereas all of the elements from the CHO2 promoter tested were active. These results are discussed in light of the differential regulation of the two promoters.

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Biochemistry, cell biology and molecular biology of lipids ofSaccharomyces cerevisiae

Daum

Lees

Bard

et al. 1998

Yeast

579

380

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The yeast Saccharomyces cerevisiae is a powerful experimental system to study biochemical, cell biological and molecular biological aspects of lipid synthesis. Most but not all genes encoding enzymes involved in fatty acid, phospholipid, sterol or sphingolipid biosynthesis of this unicellular eukaryote have been cloned, and many gene products have been functionally characterized. Less information is available about genes and gene products governing the transport of lipids between organelles and within membranes, turnover and degradation of complex lipids, regulation of lipid biosynthesis, and linkage of lipid metabolism to other cellular processes. Here we summarize current knowledge about lipid biosynthetic pathways in S. cerevisiae and describe the characteristic features of the gene products involved. We focus on recent discoveries in these fields and address questions on the regulation of lipid synthesis, subcellular localization of lipid biosynthetic steps, cross‐talk between organelles during lipid synthesis and subcellular distribution of lipids. Finally, we discuss distinct functions of certain key lipids and their possible roles in cellular processes. © 1998 John Wiley & Sons, Ltd.

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Cis and trans regulatory elements required for regulation of theCHO1gene ofSaccharomyces cerevisiae

Cited by 63 publications

References 37 publications

Regulation of Phospholipid Synthesis in the Yeast cki1Δ eki1Δ Mutant Defective in the Kennedy Pathway

Regulation of Phospholipid Synthesis in the Yeast cki1Δ eki1Δ Mutant Defective in the Kennedy Pathway

Functional characterization of the repeated UASINO element in the promoters of the INO1 and CHO2 genes of yeast

Biochemistry, cell biology and molecular biology of lipids ofSaccharomyces cerevisiae

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