Characterization of expression of the gene for human pterin carbinolamine dehydratase/dimerization cofactor of HNF1.
Pterin carbinolamine dehydratase/dimerization cofactor of HNF1 (PCD/DCoH) is a dual-function protein. In the cytoplasm it acts as a dehydratase in the regeneration of tetrahydrobiopterin, the cofactor for aromatic amino acid hydroxylases. In the nucleus, it functions as a dimerization cofactor of HNF1 and increases the transcriptional activity of HNF1. To deepen our understanding of this protein, we characterized its expression in human tissues and cells. Human PCD/DCoH was present predominantly in liver and kidney, with significant amounts in testis and ovary, trace amounts in lung, and undetectable levels in whole brain, heart, and spleen. It was expressed in all of the cells that were examined. Importantly, it was also present in the nucleus of HeLa cells, which lack HNF1, and in the cytoplasm of fibroblasts that have little or no tetrahydrobiopterin. The expression of human PCD/DCoH in the liver and nonhepatic cells was compared at both the mRNA and protein levels. Although the mRNA level in liver was only fourfold higher than that in keratinocytes and fibroblasts, the hepatic PCD/DCoH protein level was 20-fold higher than that in normal human epidermal keratinocytes and dermal fibroblasts. Cloning of the 5' and 3' untranslated region (UTR) of human keratinocyte PCD/DCoH revealed that it has 53 bp more of GC-rich 5' untranslated sequence than the published liver PCD/DCoH. In vitro transcription and translation analysis showed that the longer 5' UTR resulted in about a 35% decrease in translation efficiency. These data show that human PCD/DCoH is not only present in cells where tetrahydrobiopterin is synthesized or HNF1 is present but is a widely distributed protein. Its differential expression in different tissues and cells is regulated not only at the transcriptional level but also at the translational level.
Lei
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Laboratory of Neurochemistry, National Institute of Mental Health, Bethesda, Maryland 20892, USA.
Kaufman
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D020413:3' Untranslated Regions / Q000235:genetics; D020121:5' Untranslated Regions / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000201:enzymology; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic; D006367:HeLa Cells; D006801:Humans; D006836:Hydro-Lyases / Q000235:genetics* / Q000378:metabolism; D007962:Leukocytes / Q000166:cytology / Q000201:enzymology; D008099:Liver / Q000166:cytology / Q000201:enzymology; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D012333:RNA, Messenger; D014157:Transcription Factors; D006836:Hydro-Lyases; C076392:pterin-4a-carbinolamine dehydratase