Further characterization of a Chinese hamster ovary cell mutant requiring cholesterol and unsaturated fatty acid for growth

Chin, Jean; Chang, Ta Yuan

doi:10.1021/bi00256a025

Cited by 20 publications

(11 citation statements)

References 37 publications

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“…Studies using the CHO cell line confirm this view. SCD activity was shown to increase when CHO cells were incubated with lipid-depleted media; however, when the medium was supplemented with cholesterol, the desaturase activity was found to be reduced [50].…”

Section: Discussionmentioning

confidence: 94%

Metabolomic Analysis of SCD during Goose Follicular Development: Implications for Lipid Metabolism

Yuan

et al. 2020

Genes

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Stearoyl-CoA desaturase (SCD) is known to be an important rate-limiting enzyme in the production of monounsaturated fatty acids (MUFAs). However, the role of this enzyme in goose follicular development is poorly understood. To investigate the metabolic mechanism of SCD during goose follicular development, we observed its expression patterns in vivo and in vitro using quantitative reverse-transcription (qRT)-PCR. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to determine a cellular model of SCD function in granulosa cells (GCs) via SCD overexpression and knockdown. qRT-PCR analysis showed that SCD was abundantly expressed in the GC layer, and was upregulated in preovulatory follicles. Peak expression was found in F1 and prehierarchal follicles with diameters of 4–6 mm and 8–10 mm, respectively. We further found that mRNA expression and corresponding enzyme activity occur in a time-dependent oscillation pattern in vitro, beginning on the first day of GC culture. By LC-MS/MS, we identified numerous changes in metabolite activation and developed an overview of multiple metabolic pathways, 10 of which were associated with lipid metabolism and enriched in both the overexpressed and knockdown groups. Finally, we confirmed cholesterol and pantothenol or pantothenate as potential metabolite biomarkers to study SCD-related lipid metabolism in goose GCs.

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Section: Discussionmentioning

confidence: 94%

Metabolomic Analysis of SCD during Goose Follicular Development: Implications for Lipid Metabolism

Yuan

et al. 2020

Genes

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“…They noted that the increase in stearoyl-CoA desaturase activity was prevented when the medium was supplemented with medium that contained cholesterol (33). Subsequently, Chang and co-workers (17) isolated a mutant Chinese hamster ovary cell that both failed to induce cholesterol biosynthetic enzymes and the LDL receptor in response to cellular sterol depletion and was auxotrophic for both cholesterol and unsaturated fatty acids.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional Activation of the Stearoyl-CoA Desaturase 2 Gene by Sterol Regulatory Element-binding Protein/Adipocyte Determination and Differentiation Factor 1

Tabor¹,

Kim²,

Spiegelman³

et al. 1998

Journal of Biological Chemistry

102

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To identify genes that are transcriptionally activated by sterol regulatory element-binding proteins (SREBPs), we utilized mRNA differential display and mutant cells that express either high or low levels of transcriptionally active SREBP. This approach identified stearoyl-CoA desaturase 2 (SCD2) as a new SREBP-regulated gene.Cells were transiently transfected with reporter genes under the control of different fragments of the mouse SCD2 promoter. Constructs containing >199 base pairs of the SCD2 proximal promoter were activated following incubation of cells in sterol-depleted medium or as a result of co-expression of SREBP-1a, SREBP-2, or rat adipocyte determination and differentiation factor 1 (ADD1). Electromobility shift assays and DNase I footprint analysis demonstrated that recombinant SREBP-1a bound to a novel cis element (5-AGCAGATTGTG-3) in the proximal promoter of the SCD2 gene. The finding that the endogenous SCD2 mRNA levels were induced when wild-type Chinese hamster ovary fibroblasts were incubated in sterol-deficient medium is consistent with a role for SREBP in regulating transcription of the gene.These studies identify SCD2 as a new member of the family of genes that are transcriptionally regulated in response to changing levels of nuclear SREBP/ADD1. In addition, the sterol regulatory element in the SCD2 promoter is distinct from all previously characterized motifs that confer SREBP-and ADD1-dependent transcriptional activation.

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“…LXR ␣ is the oxysterol receptor that mediates regulation of gene expression by cholesterol or its metabolites (73,74). Mice lacking LXR ␣ receptor expressed low levels of SCD mRNA in the presence or absence of cholesterol feeding while the wild-type animals fed 2% cholesterol had their SCD mRNA levels up-regulated 4-fold (72). As expected, the mRNA levels corresponding to several enzymes involved in cholesterol biosynthesis were up-regulated in the LXR ␣ knockout animals and down-regulated in the wild-type animals upon cholesterol feeding.…”

Section: Cholesterol As a Regulator Of Scd Gene Expressionmentioning

confidence: 99%

Regulation of stearoyl-CoA desaturase by polyunsaturated fatty acids and cholesterol

Ntambi

1999

Journal of Lipid Research

639

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The lipid composition of cellular membranes is regulated to maintain membrane fluidity. A key enzyme involved in this process is the membrane-bound stearoyl-CoA desaturase (SCD) which is the rate-limiting enzyme in the cellular synthesis of monounsaturated fatty acids from saturated fatty acids. A proper ratio of saturated to monounsaturated fatty acids contributes to membrane fluidity. Alterations in this ratio have been implicated in various disease states including cardiovascular disease, obesity, non-insulindependent diabetes mellitus, hypertension, neurological diseases, immune disorders, and cancer. The regulation of stearoyl-CoA desaturase is therefore of considerable physiological importance and its activity is sensitive to dietary changes, hormonal imbalance, developmental processes, temperature changes, metals, alcohol, peroxisomal proliferators, and phenolic compounds. Two mouse and rat SCD genes (SCD1 and SCD2) and a single human SCD gene have been cloned and characterized. In the past several years we have studied the dietary influences on the genetic expression of the mouse stearoyl-CoA desaturase. The expression of the mouse SCD genes is regulated by polyunsaturated fatty acids and cholesterol at the levels of transcription and mRNA stability. Promoter elements that are responsible for the polyunsaturated fatty acid repression colocalize with the promoter elements for SREBP-mediated regulation of the SCD genes.It is the goal of this review to provide an overview of the genetic regulation of the stearoyl-CoA desaturase in response to dietary polyunsaturated fatty acids and cholesterol. -Ntambi, J. M. Regulation of stearoyl-CoA desaturase by polyunsaturated fatty acids and cholesterol.

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Further characterization of a Chinese hamster ovary cell mutant requiring cholesterol and unsaturated fatty acid for growth

Cited by 20 publications

References 37 publications

Metabolomic Analysis of SCD during Goose Follicular Development: Implications for Lipid Metabolism

Metabolomic Analysis of SCD during Goose Follicular Development: Implications for Lipid Metabolism

Transcriptional Activation of the Stearoyl-CoA Desaturase 2 Gene by Sterol Regulatory Element-binding Protein/Adipocyte Determination and Differentiation Factor 1

Regulation of stearoyl-CoA desaturase by polyunsaturated fatty acids and cholesterol

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