miR-584 mediates post-transcriptional expression of lactoferrin receptor in Caco-2 cells and in mouse small intestine during the perinatal period
Copyright policy )miR-584 mediates post-transcriptional expression of lactoferrin receptor in Caco-2 cells and in mouse small intestine during the perinatal period
MicroRNAs function as gene expression modulators that are critical for mammalian development. Lactoferrin receptor on the apical membrane of enterocytes has been suggested to play key roles in the absorption of lactoferrin-bound iron from breast milk. The objective of this study was to identify mechanisms of microRNA mediated post-transcriptional regulation of the lactoferrin receptor. Sequence analyses revealed that the miR-584 sequence is identical in human, mouse and rat, and there is a conserved region complementary to the seed region (5' nucleotides 2-8) of miR-584 within the lactoferrin receptor mRNA-3'-untranslated region. miR-584 was further found to co-localize with lactoferrin receptor mRNA in mouse small intestine. The 3'-untranslated region of human lactoferrin receptor mRNA was cloned into pGL3-control luciferase reporter vector. By luciferase reporter assays in HEK293 cells, miR-584 mimic specifically repressed the reporter activity in a dose-dependent manner. miR-584 mimic reduced endogenous lactoferrin receptor protein expression in Caco-2 cells, without significantly affecting the mRNA level. We also determined that miR-584 expression is inversely correlated with lactoferrin receptor mRNA and protein expression. Taken together, we propose that miR-584 contributes to the post-transcriptional expression of lactoferrin receptor during the perinatal period. These findings demonstrate a novel example of how microRNAs may be involved in regulation of nutrient metabolism in the newborn.
- Australian National University Australia
- University of California, Davis United States
molecular cloning, 570, MiR-584, Molecular Sequence Data, Down-Regulation, Receptors, Cell Surface, animal cell, RNA Transport, Mice, Species Specificity, cell strain CACO 2, Intestine, Small, Animals, Humans, human, RNA, Messenger, fluorescence in situ hybridization, 3' Untranslated Regions, cell strain HEK293, mouse, Conserved Sequence, Keywords: lactoferrin, cell culture, Binding Sites, microRNA, Base Sequence, Microvilli, human cell, article, Gene Expression Regulation, Developmental, luciferase, n Lactoferrin receptor (LfR), Gene Expression Regulation, Neoplastic, MicroRNAs, Animals, Newborn, concentration response, genetic transfection, MicroRNA (miRNA), Caco-2 Cells, immunoblotting
molecular cloning, 570, MiR-584, Molecular Sequence Data, Down-Regulation, Receptors, Cell Surface, animal cell, RNA Transport, Mice, Species Specificity, cell strain CACO 2, Intestine, Small, Animals, Humans, human, RNA, Messenger, fluorescence in situ hybridization, 3' Untranslated Regions, cell strain HEK293, mouse, Conserved Sequence, Keywords: lactoferrin, cell culture, Binding Sites, microRNA, Base Sequence, Microvilli, human cell, article, Gene Expression Regulation, Developmental, luciferase, n Lactoferrin receptor (LfR), Gene Expression Regulation, Neoplastic, MicroRNAs, Animals, Newborn, concentration response, genetic transfection, MicroRNA (miRNA), Caco-2 Cells, immunoblotting
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