Stearoyl-CoA desaturase gene expression in lymphocytes.

B lymphocytes from the spleens of normal (BALB/c) and autoimmune (MRL/lpr) strains of mice express the SCD-2 form of stearoyl-CoA desaturase as opposed to the SCD-1 form of the gene which is expressed in liver. However, whereas BALB/c T cells did not express SCD-1 or SCD-2, both BALB/c thymocytes and MRL/lpr T cells expressed SCD-2, suggesting a developmental down-regulation of SCD-2 within the T cell lineage. Northern analyses also revealed the expression of SCD-2 in the T cell lines BW5147, CTLL-2 and HT-2 and in BCL1, a B cell line. SCD-1 expression was not detected in any of the lymphoid cells tested. Finally, we show that SCD-2 gene expression is inhibited by arachidonic acid (20:4). These results demonstrate the complexity of SCD-2 regulation in lymphoid cells.     

The OLE1 gene of Saccharomyces cerevisiae encodes the delta 9 fatty acid desaturase and can be functionally replaced by the rat stearoyl-CoA desaturase gene

Strains of Saccharomyces cerevisiae bearing the ole1 mutation are defective in unsaturated fatty acid (UFA) synthesis and require UFAs for growth. A previously isolated yeast genomic fragment complementing the ole1 mutation has been sequenced and determined to encode the delta 9 fatty acid desaturase enzyme by comparison of primary amino acid sequence to the rat liver stearoyl-CoA desaturase. The OLE1 structural gene encodes a protein of 510 amino acids (251 hydrophobic) having an approximate molecular mass of 57.4 kDa. A 257-amino acid internal region of the yeast open reading frame aligns with and shows 36% identity and 60% similarity to the rat liver stearoyl-CoA desaturase protein. This comparison disclosed three short regions of high consecutive amino acid identity (greater than 70%) including one 11 of 12 perfect residue match. The predicted yeast enzyme contains at least four potential membrane-spanning regions and several shorter hydrophobic regions that align exactly with similar sequences in the rat liver protein. An ole1 gene-disrupted yeast strain was transformed with a yeast-rat chimeric gene consisting of the promoter region and N-terminal 27 codons of OLE1 fused to the rat desaturase coding sequence. Fusion gene transformants displayed near equivalent growth rates and modest lipid composition changes relative to wild type yeast control implying a significant conservation of delta 9 desaturase tertiary structure and efficient interaction between the rat desaturase and yeast cytochrome b5.     

低温和外源不饱和脂肪酸对高山被孢霉脂肪酸脱氢酶基因表达的影响

[目的]在转录水平上研究低温和外源不饱和脂肪酸对于高山被孢霉脂肪酸脱氢酶基因的表达调控机制.[方法]通过实时定量PCR技术和启动子报告基因融合载体的方法,研究低温和外源不饱和脂肪酸对于高山被孢霉3种脂肪酸脱氢酶基因表达随时间进程的影响.[结果]实时定量PCR的结果表明:低温对于3种脂肪酸脱氢酶基因的转录具有激活作用,外源不饱和脂肪酸对基因转录起抑制作用,而且这两种作用都是快速响应的,随时间延长逐渐减弱并消失.脂肪酸组成测定结果证明了基因转录水平变化与对应产物变化之间没有相关性.低温能够在短时间内诱导pFAD6启动子活性增加,并随时间延长而持续增强 ;外源不饱和脂肪酸对pFAD6启动子活性起抑制作用,其不饱和度和浓度越高,抑制作用越强,而且抑制作用是快速且持续的.[结论]低温和外源不饱和脂肪酸除了在转录水平上调控高山被孢霉脂肪酸脱氢酶基因表达发生变化之外,可能主要在转录后水平上介导了胞内脂肪酸组成的变化.而且,脂肪酸脱氢酶基因的表达可能受到胞内脂肪酸组成变化的反馈调节作用.本文首次在转录水平上对高山被孢霉脂肪酸脱氢酶基因的表达调控机制进行了探索,为深入了解脂肪酸脱氢酶基因表达及多不饱和脂肪酸合成对外界信号的应答机制提供了有用信息,也对应用微生物发酵和转基因技术生产不饱和脂肪酸具有指导意义.     

The aging human orbitofrontal cortex: decreasing polyunsaturated fatty acid composition and associated increases in lipogenic gene expression and stearoyl-CoA desaturase activity

Orbitofrontal cortex (OFC, Brodmann area 10) gray matter volume reductions and selective reductions in docosahexaenoic acid (DHA, 22:6n-3) are observed in adult patients with major depressive disorder (MDD). OFC gray matter volume also decreases with advancing age in healthy subjects. To examine if OFC gray matter DHA composition decreases during normal aging, we determined age-related changes in OFC gray matter fatty acid composition by gas chromatography in subjects aged 29-80 years (n=30). We additionally determined elongase (HELO1), delta-5 desaturase (FASD1), delta-6 desaturase (FASD2), peroxisomal (PEX19), and stearoyl-CoA desaturase (SCD) mRNA expression in the same tissues. Increasing age was associated with a progressive decline in polyunsaturated fatty acid (PUFA) composition, including DHA and arachidonic acid (AA, 20:4n-6), and transient, apparently compensatory, elevations in elongase and desaturase gene expression. The age-related reduction in PUFA composition was inversely correlated with SCD expression and activity resulting in elevations in monounsaturated fatty acid composition. These dynamic age-related changes in OFC gray matter fatty acid composition and biosynthetic gene expression may contribute to the progressive decline in OFC gray matter volume found with advancing age. The implications of age-related reductions in OFC PUFA composition for affective dysregulation in the elderly are discussed.     

Arachidonic acid suppression of fatty acid synthase gene expression in cultured rat hepatocytes

Rat hepatocytes were maintained in a serum-free, hormonally defined medium supplemented with 50-500 microM albumin-bound 20:1 (n-9) vs 20:4 (n-6). The induction of fatty acid synthase mRNA by a mix of insulin/dexamethasone/T3 was inhibited in a dose dependent fashion by 20:4 (n-6). The abundance of beta-actin mRNA was not suppressed by 20:4 (n-6). The expression of fatty acid synthase was actually stimulated 2-fold by 20:1 (n-9). It would appear that the in vivo inhibition of fatty acid synthase gene expression by dietary polyunsaturated fatty acids is a specific hepatocelluar event.     

Increased expression of the gene encoding stearoyl-CoA desaturase 1 in human bladder cancer

Bladder cancer is a common disease and a significant cause of death worldwide. There is thus great interest in identifying a diagnostic and prognostic biomarker, as well as gaining an understanding of the molecular basis of bladder cancer. Stearoyl-CoA desaturase 1 gene (SCD1) is highly overexpressed in many human cancers. However, the expression of SCD1 has not yet been investigated in patients with bladder cancer. Here, we document that (a) the SCD1 is highly overexpressed in human bladder cancer; (b) high expression of SCD1 is more frequently observed in the late stage of disease and patients with lymph node metastasis; (c) bladder cancer patients with a higher SCD1 mRNA level have a poorer survival rate than those with normal SCD1 expression. Overall, this is the first report to indicate an association between SCD1 mRNA level and clinical indicators of human bladder cancer. Our study has provided evidence supporting the potential role of SCD1 as a biomarker for human bladder cancer prognosis.     

Arachidonic acid regulates surface expression of epithelial sodium channels

Epithelial Na+ channels (ENaCs) are regulated by the phospholipase A2 (PLA2) product arachidonic acid. Pharmacological inhibition of PLA2 with aristolochic acid induced a significant increase in amiloride-sensitive currents in Xenopus oocytes expressing ENaC. Arachidonic acid or 5,8,11,14-eicosatetraynoic acid (ETYA), a non-metabolized analog of arachidonic acid, induced a time-dependent inhibition of Na+ transport. These effects were also observed by co-expression of a calcium-independent or a calcium-dependent PLA2. Channels with a truncated alpha, beta,or gamma C terminus were not inhibited by arachidonic acid or ETYA. Furthermore, mutation of Tyr618 in the PY motif of the beta subunit abrogated the inhibitory effect of ETYA, suggesting that intact PY motifs participate in arachidonic acid-mediated ENaC inhibition. Analyses of channels expressing a series of beta subunit C-terminal truncations revealed a second region N-terminal to the PY motif (spanning residues betaVal580-betaGly599) that allowed for ETYA-mediated ENaC inhibition. Analyses of both ENaC surface expression and ENaC trafficking with mutants that either gate channels open or closed in response to [(2-(trimethylammonium) ethyl] methanethiosulfonate bromide, or with brefeldin A, suggest that ETYA reduces channel surface expression by inhibiting ENaC exocytosis and increasing ENaC endocytosis.