A Non-Synonymous Single Nucleotide Polymorphism in FASN Gene Alters FASN Enzyme Activity in Subcutaneous and Intramuscular Adipose Tissue in Holstein Friesian Steers

Cancino-Baier, David; Muñoz, Erwin; Quiñones, John; Beltrán, Jorge F.; Fuentes, Fernanda; Farías, Jorge; Lorenzo, José Manuel; Diaz, Rommy; Inostroza, Karla; Ferraz, José Bento Sterman; Sepúlveda, Néstor

doi:10.2478/aoas-2020-0059

Abstract

The FASN enzyme plays a key role in fatty acids synthesis as the main long-chain fatty acid synthesizer. A non-synonymous SNP (single nucleotide polymorphism) (g.17925A>G) located in the thioesterase domain of this enzyme and an effect in fat deposition has been observed, but has not been evaluated in this breed and, moreover, the reason whereby this occurs remains unclear. The objective of this study was to evaluate the effect of this SNP on the activity of FASN enzyme in subcutaneous and intramuscular adipose tissue from Holstein Friesian steers. To achieve this, 196 animals were sampled in a local abattoir, genotyped for the FASN g.17924A>G SNP and characterized for fatty acid profile. Then a sub-sample of 20 animals per genotype were selected to extract the total protein from subcutaneous and intramuscular adipose tissue to estimate the FASN enzyme activity. The FASN activity for each genotyped animal was assessed indirectly by measuring the decrease in the absorbance of NADPH at 340 nm by spectrophotometry in a 24 well plate in the presence of Acetyl-CoA, Malonyl-CoA, and NADPH. To assess the impact of SNP induced amino acid changes in FASN protein structure, in-silico simulations were performed. Our results indicated that FASN g.17924A>G SNP induces a change in the enzyme activity in subcutaneous adipose tissue, which is higher when the AA genotype is present and lower in the presence of the AG genotype. The in-silico analysis of the amino acid substitution shows that there was a structural change in the dimeric form of the protein between genotypes. Moreover, the global energy between subunits is lower and more favorable when the AA genotype is present and higher and less favorable for the AG genotype. It was also found that the fatty acid profile of subcutaneous adipose tissue was affected when the AG genotype was present, decreasing the C16:0 fatty acid levels and increasing the C18:0 fatty acid levels. The FASN g.17924A>G SNP alters the FASN enzyme structure and activity, leading to a variation in the fatty acid composition of subcutaneous adipose tissue in Holstein Friesian steers. Implications: This SNP could be considered as a tool to improve the fat deposition or marbling and the fatty acid profile in cattle.

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A Non-Synonymous Single Nucleotide Polymorphism in FASN Gene Alters FASN Enzyme Activity in Subcutaneous and Intramuscular Adipose Tissue in Holstein Friesian Steers

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