The Se-dependent expression of two selenoproteins, cytosolic glutathione peroxidase (cGPx) and type I iodothyronine-5′-deiodinase (5′DI), was investigated in the porcine epithelial kidney cell line LLC-PK1 in serum-free medium. The selenite-dependent expression of cGPx and 5′DI was revealed by enzyme-activity measurements, affinity labelling of 5′DI, metabolic labelling of proteins with 75Se and steady-state mRNA analysis. The expression of the two enzymes strongly depended on selenite concentrations of the culture medium. cGPx required 2-fold higher selenite levels than 5′DI to reach half-maximal activity. The Se-dependent enzyme activities were approximately paralleled by the corresponding steady-state mRNA levels. The response of the two enzymes to Se supply was further characterized by kinetic Se-depletion and -repletion experiments. Upon removal of medium selenite, cGPx activity decreased exponentially, whereas after an initial decrease over 1-2 days, 5′DI levels completely recovered during a further 2 days. These data indicate a differential Se-dependent regulation of the two selenoproteins, with 5′DI being preferentially supplied with the trace element Se, thus ensuring a continuous cellular capacity for thyroid-hormone activation, even under Se-deficient conditions. The abundant cGPx in cells with sufficient Se supply might serve as a cellular Se store which can be mobilized for the synthesis of more vital selenoproteins such as 5′DI under shortage conditions. Thus, a cellular hierarchy of selenoprotein expression, reflected by different individual regulation mechanisms at the transcriptional and post-transcriptional level, adds to the previously recognized tissue-specific hierarchy of Se retention.
Differential selenium-dependent expression of type I 5′-deiodinase and glutathione peroxidase in the porcine epithelial kidney cell line LLC-PK1
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M Gross, M Oertel, J Köhrle; Differential selenium-dependent expression of type I 5′-deiodinase and glutathione peroxidase in the porcine epithelial kidney cell line LLC-PK1. Biochem J 15 March 1995; 306 (3): 851–856. doi: https://doi.org/10.1042/bj3060851
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