RESUMO
Carotenoids are currently investigated regarding their potential to lower the risk of chronic disease and to combat vitamin A deficiency. Surprisingly, responses to dietary supplementation with these compounds are quite variable between individuals. Genome-wide studies have associated common genetic polymorphisms in the BCO1 gene with this variability. The BCO1 gene encodes an enzyme that is expressed in the intestine and converts provitamin A carotenoids to vitamin A-aldehyde. However, it is not clear how this enzyme can impact the bioavailability and metabolism of other carotenoids such as xanthophyll. We here provide evidence that BCO1 is a key component of a regulatory network that controls the absorption of carotenoids and fat-soluble vitamins. In this process, conversion of ß-carotene to vitamin A by BCO1 induces via retinoid signaling the expression of the intestinal homeobox transcription factor ISX. Subsequently, ISX binds to conserved DNA-binding motifs upstream of the BCO1 and SCARB1 genes. SCARB1 encodes a membrane protein that facilitates absorption of fat-soluble vitamins and carotenoids. In keeping with its role as a transcriptional repressor, SCARB1 protein levels are significantly increased in the intestine of ISX-deficient mice. This increase results in augmented absorption and tissue accumulation of xanthophyll carotenoids and tocopherols. Our study shows that fat-soluble vitamin and carotenoid absorption is controlled by a BCO1-dependent negative feedback regulation. Thus, our findings provide a molecular framework for the controversial relationship between genetics and fat-soluble vitamin status in the human population.
Assuntos
Xantofilas/metabolismo , Zeaxantinas/metabolismo , Animais , Sequência de Bases , Sítios de Ligação , Olho/metabolismo , Feminino , Expressão Gênica , Absorção Intestinal , Masculino , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Regiões Promotoras Genéticas , Receptores Depuradores Classe B/fisiologia , Solubilidade , Fatores de Transcrição/fisiologia , Ativação TranscricionalRESUMO
The scavenger receptor class B, type I (SR-BI) is an important player in regulation of mammalian lipid homeostasis. We therefore wanted to study this receptor in Atlantic salmon (Salmo salar L.), which requires a diet with particular high lipid content. We have for the first time cloned and characterized SR-BI from a salmonid fish. The predicted 494 amino acid protein contained two transmembrane domains, several putative N-glycosylation sites, and showed 72% sequence identity with the predicted homolog from zebrafish. SR-BI expression was analyzed by reverse transcription Real-Time PCR in several tissues, and a high relative expression in salmon midgut was detected, which may suggest that SR-BI has a role in uptake of lipids from the diet. We also expressed a construct of salmon myc-tagged SR-BI in salmon TO cells and HeLa cells, which gave a protein of approximately 80 kDa on reducing SDS-PAGE using an antibody against the myc-epitope. Immunofluorescence microscopy analyses of the salmon SR-BI protein in transiently transfected HeLa cells revealed staining in the cell periphery and in some intracellular membranes, but not in the nucleus, which indicated that the salmon protein may be a functional membrane protein. We also observed a high degree of co-localization using an anti-peptide SR-BI antiserum. We found that 20 microg mL(-1) insulin up-regulated the SR-BI mRNA levels in primary cultures of salmon hepatocytes relative to untreated cells. Oleic acid, EPA, DHA, or dexamethasone did not affect the relative expression of SR-BI in this liver model system. In conclusion, the salmon SR-BI cDNA encoded a protein with several features common to those of mammalian species. SR-BI gene expression was high in the intestine, which leads us to propose that SR-BI may contribute to the uptake of lipids from the diet.