Differential selenium-dependent expression of type I 5'-deiodinase and glutathione peroxidase in the porcine epithelial kidney cell line LLC-PK1.
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.
Gross
M
M
Max Planck Institut für Experimentelle Endokrinologie, Hannover, Federal Republic of Germany.
Oertel
M
M
Köhrle
J
J
D000818:Animals; D003470:Culture Media; D004848:Epithelium / Q000201:enzymology; D005979:Glutathione Peroxidase / Q000096:biosynthesis*; D007453:Iodide Peroxidase / Q000096:biosynthesis*; D018374:LLC-PK1 Cells; D012643:Selenium / Q000494:pharmacology*; D013552:Swine
D003470:Culture Media; D007453:Iodide Peroxidase; D005979:Glutathione Peroxidase; D012643:Selenium