Abstract
Biotin is covalently attached to carboxylases and histones in a reaction catalysed by holocarboxylase synthetase (HCS). Here we determined whether biotin concentrations in culture media affect the expression of genes maintaining biotin homeostasis in human lymphoid (Jurkat) cells. As a secondary objective we determined whether mechanisms of biotin homeostasis are distinct in cells with great metabolic activities, i.e., human hepatocarcinoma (HepG2) cells. Jurkat cells were cultured in media containing deficient, physiological, or high concentrations of biotin for 36 d. While the abundances of biotinylated carboxylases responded rapidly (<3 d) to altered biotin supply, changes in histone biotinylation were not apparent until day 36 when biotin supplementation caused increased biotinylation of histones in nuclear extracts. The expression of genes linked with biotin homeostasis depended on biotin in Jurkat cells. The expression of biotin transporters SMVT and MCT1 increased in biotin‐deficient cells, whereas the expression of HCS increased in biotin‐supplemented cells. In ongoing studies we investigate the effects of biotin supply on histone biotinylation at distinct genomic loci by ChIP assays, and we compare biotin homeostasis in Jurkat cells to HepG2 liver cells. (Supported by UNL ARD Hatch Act funds, NIH DK063945, DK077816 and ES015206, USDA CSREES 2006‐35200‐17138, and NSF EPSCoR EPS‐0701892.)
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