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Role of phospholipase D in salycilic acid signaling in Arabidopsis thaliana suspension cells

ABSTRACT: Our goal is to identify Salycilic Acid responsive genes dependent on PLD activation. - The experiment is done on Arabidopsis suspension cells, ecotype Columbia : Research of phospholipase D (PLD) activity implication in the response to SA. The use of primary alcohol, like N-butanol, makes possible to derive PLD activity towards the production of phosphatidylalcohol with the detriment of phosphatidic acid, the product of the PLD. Thus, in the presence of n-butanol, the response of the genes to SA dependent on phosphatidic acid will see their response disturbed. On the contrary, in the presence of tertiary butanol, the response of the genes should not be disturbed, tertiary alcohols not acting on the PLD. Keywords: treated vs untreated comparison

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE9695 | GEO | 2007/11/29

SECONDARY ACCESSION(S): PRJNA103597

REPOSITORIES: GEO

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Project description:A time course of the effect of overexpressing GPI-PLD on global gene expression in HepG2 cells was compared to control virus. Experiment Overall Design: This experiment compares the global gene expression in HepG2 cells that were either transduced with an adenovirus expressing GPI-phospholipase D or an adenovirus expressing beta galactosidase as a control. Treatment was for 6 or 12 hrs. Four replicates for each condition were used. 10 micrograms of total RNA was assayed per genechip using standard Affymetrix protocols. CONTEXT: Recent studies demonstrated that de novo lipogenesis is increased in patients with nonalcoholic fatty liver disease (NAFLD). Patients with NAFLD also have plasma lipid abnormalities. These lipid abnormalities may in part be related to insulin resistance, which is common in patients with NAFLD. Insulin resistance is associated with alterations in proteins involved in lipid metabolism including glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD), which is involved in triglyceride metabolism. OBJECTIVE: The objective of the study was to determine whether alterations in serum and hepatic levels of GPI-PLD occur in patients with NAFLD. DESIGN AND PATIENTS: We examined the following: 1) levels of serum GPI-PLD in nondiabetics with nonalcoholic steatohepatitis, compared with matched controls; 2) hepatic expression of GPI-PLD mRNA in patients with normal liver or NAFLD; and 3) effect of overexpressing GPI-PLD vs. beta-galactosidase (control) on global gene expression in a human hepatoma cell line. RESULTS: The serum levels of GPI-PLD were significantly higher in patients with nonalcoholic steatohepatitis than in matched controls (119 +/- 24 vs.105 +/- 15 microg/ml, P = 0.047). The hepatic expression of GPI-PLD mRNA was increased nearly 3-fold in NAFLD patients, compared with patients with normal liver (3.1 +/- 2.6 vs. 1.1 +/- 1.0 arbitrary units per microgram total RNA, P = 0.026). Finally, overexpressing GPI-PLD was associated with an increase in de novo lipogenesis genes. CONCLUSIONS: Patients with NAFLD have elevated serum levels and hepatic expression of GPI-PLD, and its overexpression in vitro is associated with increased expression of de novo lipogenesis genes. These results suggest that GPI-PLD may play a role in the pathogenesis of NAFLD and/or its metabolic features and warrants further investigation.

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Role of phospholipase D in salycilic acid signaling in Arabidopsis thaliana suspension cells

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