The C-terminal region of cis-retinol/androgen dehydrogenase 1 (CRAD1) confers ER localization and in vivo enzymatic function.
Retinoic acid is generated from retinol (vitamin A) by the sequential actions of two different classes of enzymes, retinol dehydrogenases and retinal dehydrogenases. Several enzymes implicated in this process have been identified and characterized in vitro. However, our understanding of the cell biological function and regulation of this process is limited. To get further knowledge regarding the regulation of RA biosynthesis, we have determined possible regulatory mechanisms at the transcriptional and post-transcriptional levels for the prototypic microsomal retinol dehydrogenase cis-retinol/androgen dehydrogenase 1 (CRAD1). We note that the expression and stability of the enzyme are only moderately controlled by the retinoid status. Instead, we find that the cytosolic tail dramatically affects the activity of the enzyme, and we have mapped the structural elements required for ER retention and in vivo functional activity, respectively. Although inactive tail-deletion mutants display an abnormal subcellular localization, restoration of ER localization per se is not sufficient for enzymatic activity suggesting that additional trans-acting components interacting with, or modifying, the cytosolic tail are required for controlling the activity of the enzyme in vivo.
Lidén
Martin
M
Ludwig Institute for Cancer Research, Stockholm Branch, Box 240, S-17177 Stockholm, Sweden.
Tryggvason
Kristian
K
Eriksson
Ulf
U
D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D017384:Sequence Deletion; D014158:Transcription, Genetic
D007136:Immunoglobulins; D008565:Membrane Proteins; D000429:Alcohol Oxidoreductases; C109795:cis-retinol - androgen dehydrogenase