Molecular cloning and sequence of a cholesterol-repressible enzyme related to prenyltransferase in the isoprene biosynthetic pathway.

Clarke, Catherine F.; Tanaka, Richard D.; Svenson, Karen L.; Wamsley, M; Fogelman, Alan M.; Edwards, Peter A.

doi:10.1128/mcb.7.9.3138

Cited by 122 publications

(64 citation statements)

References 65 publications

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“…which is similar to the molecular masses of other FPP synthases (40, 39, and 43 kD for the yeast, rat, and plant enzymes, respectively) (Clarke et al, 1987;Anderson et al, 1989;Sheares et al, 1989;Hugueney and Camara, 1990;Teruya et al, 1990). The deduced amino acid sequence of the encoded protein has 80% identity with and 90% similarity to that of FPP synthase identified from A. thaliana (GenBank accession No.…”

supporting

confidence: 50%

A cDNA Encoding Farnesyl Pyrophosphate Synthase in White Lupin

Attucci

Aitken²,

Ibrahim³

et al. 1995

Plant Physiol.

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supporting

confidence: 50%

A cDNA Encoding Farnesyl Pyrophosphate Synthase in White Lupin

Attucci

Aitken²,

Ibrahim³

et al. 1995

Plant Physiol.

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“…For example, a number of other enzymes involved in cholesterol biosynthesis are regulated by sterols. These enzymes include mevalonate kinase (39), farnesyl-pyrophosphate synthase (9), and squalene synthase (13). If regulation of these enzymes is defective in SRD-1, SRD-2, and SRD-6 cells, then it is likely that these enzymes are regulated by the same mechanism that controls HMG-CoA reductase, HMG-CoA synthase, and the LDL receptor.…”

Section: Resultsmentioning

confidence: 99%

Loss of transcriptional activation of three sterol-regulated genes in mutant hamster cells.

Evans¹,

Metherall²

1993

Mol. Cell. Biol.

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Cholesterol biosynthesis and uptake are controlled by a classic end product-feedback mechanism whereby elevated cellular sterol levels suppress transcription of the genes encoding 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase, HMG-CoA reductase, and the low-density lipoprotein receptor. The 5'-flanking region of each gene contains a common cis-acting element, designated the sterol regulatory element (SRE) cells is limited to transcriptional regulation, since posttranscriptional mechanisms of sterol-mediated regulation were intact: the cells retained the ability to posttranscriptionally suppress HMG-CoA reductase activity and to stimulate acyl-CoA:cholesterol acyltransferase activity. These results suggest that SRD-6 cells lack a factor required for SRE-dependent transcriptional activation. We contrast these cells with a previously isolated oxysterol-resistant cell line (SRD-2) that lacks a factor required for SRE-dependent transcriptional suppression and propose a model for the role of these genetically defined factors in sterol-mediated transcriptional regulation.Mammalian cells demonstrate exquisite control over the level of free cholesterol within the cell (reviewed in reference 17). This control is achieved through the concerted action of a number of mechanisms, including the coordinate transcriptional suppression of the genes encoding 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase, HMG-CoA reductase, and the low-density lipoprotein (LDL) receptor. HMG-CoA synthase and HMG-CoA reductase are the first two committed steps in the pathway of cholesterol biosynthesis, and transcriptional suppression of these genes decreases the rate of cholesterol synthesis. The LDL receptor mediates endocytosis of cholesterol-rich lipoprotein particles such as LDL, and transcriptional suppression of the LDL receptor results in a decreased rate of cholesterol uptake. Transcriptional suppression of these three genes appears to be mediated through a common cis-acting sequence, termed the sterol regulatory element (SRE), that resides in the 5'-flanking region of each gene. A protein that interacts with the SRE of the LDL receptor promoter has recently been identified and purified (1,40 levels rise, the rate of degradation of HMG-CoA reductase is enhanced. This increased degradation requires the membrane-spanning domain of the protein, which anchors it to the endoplasmic reticulum (14). This increased degradation results in decreased HMG-CoA reductase activity and a consequent decrease in the rate of cholesterol synthesis. Sterols also inhibit translation of HMG-CoA reductase mRNA (28), which further decreases the rate of cholesterol synthesis. In addition, sterols stimulate acyl-CoA:cholesterol acyltransferase (ACAT) activity by a mechanism that does not require new protein synthesis (4, 16). ACAT resides in the endoplasmic reticulum and catalyzes the transfer of long-chain fatty acyl residues from acyl-CoA to the 3-hydroxyl group of cholesterol to form cholesteryl esters. While free cholesterol can be deleter...

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“…The radiolabeled probe was used to screen -2 x 105 recombinants. Plaque hybridization was performed as described (8), after which the filters were washed at 680C with 2 x SSC (1x SSC = 0.15 M NaCI/0.015 M sodium citrate, pH 7) containing 0.1% SDS, followed by washes with 0.lx SSC containing 0.1% SDS. Fifteen cDNA clones were isolated.…”

Section: Methodsmentioning

confidence: 99%

Molecular cloning of mevalonate kinase and regulation of its mRNA levels in rat liver.

Tanaka¹,

Lee²,

Schafer³

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

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Mevalonate kinase [ATP:(R)-mevalonate 5-phosphotransferase, EC 2.7.1.36] may be a regulatory site in the cholesterol biosynthetic pathway, and a mutation in the gene coding for this enzyme is thought to cause the genetic disease mevalonic aciduria. To characterize this enzyme, a rat liver cDNA library was screened with a monospecific antibody, and a 1.7-kilobase cDNA clone coding for mevalonate kinase was isolated. A mutation in the gene coding for mevalonate kinase is presumed to be the cause of the recently discovered genetic disease mevalonic aciduria (4, 5). The genetic disease is transmitted as an autosomal recessive trait, and there are six reported cases (6). Subjects with mevalonic aciduria have extremely high levels of mevalonate in their plasma and urine, and cells from these subjects have <10% of the normal levels of mevalonate kinase activity (4-6). As a first step in identifying the molecular defect causing mevalonic aciduria and to study the regulation of mevalonate kinase activity, we have isolated and characterized a cDNA clone coding for rat mevalonate kinase.* Data are also presented to show that long-term regulation of enzyme activity in rat liver is controlled by changes in the levels of mevalonate kinase mRNA. MATERIALS AND METHODSPreparation of the Antiserum. Monospecific antisera to rat mevalonate kinase was prepared in rabbits by using the purified enzyme (3), and the IgG fraction was collected by affinity chromatography (7).Isolation of a cDNA Clone Coding for Mevalonate Kinase. A Agtll cDNA library (8) derived from mRNA purified from the livers of rats treated with diets containing 5% cholestyramine and 0.1% lovastatin was kindly provided by Peter Edwards (UCLA). Positive clones were identified by immunoscreening (9). To obtain a full-length cDNA clone, the cDNA insert from one of the clones was isolated and radiolabeled by random priming (10) to a specific activity of 109 cpm per Ag ofDNA. The radiolabeled probe was used to screen -2 x 105 recombinants. Plaque hybridization was performed as described (8), after which the filters were washed at 680C with 2 x SSC (1x SSC = 0.15 M NaCI/0.015 M sodium citrate, pH 7) containing 0.1% SDS, followed by washes with 0.lx SSC containing 0.1% SDS. Fifteen cDNA clones were isolated. The cDNA inserts were subcloned into the pGEM-3Z vector (Promega) for restriction enzyme mapping or into M13 vectors for DNA sequencing.DNA Sequencing. The DNA sequence of the cDNA was determined by the dideoxynucleotide chain-termination method (11). Sequencing reactions using the Klenow fragment of DNA polymerase I (New England Biolabs) or the modified T7 DNA polymerase (Sequenase, United States Biochemicals) were performed following the manufacturer's protocol. The DNA and protein sequences were aligned using the Intelligenetics computer program, and the EMBL/ GenBank and PIR data bases were searched for homologies to the DNA sequence and protein sequence of mevalonate kinase.Expression of the Mevalonate Kinase cDNA in Saccharomyces cerevisiae. A 1.3-kilobase...

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Molecular cloning and sequence of a cholesterol-repressible enzyme related to prenyltransferase in the isoprene biosynthetic pathway.

Cited by 122 publications

References 65 publications

A cDNA Encoding Farnesyl Pyrophosphate Synthase in White Lupin

A cDNA Encoding Farnesyl Pyrophosphate Synthase in White Lupin

Loss of transcriptional activation of three sterol-regulated genes in mutant hamster cells.

Molecular cloning of mevalonate kinase and regulation of its mRNA levels in rat liver.

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