title: MicroRNA-939 down-regulates CD2-associated protein by targeting promoter in HEK-293T cells.
abstract: CD2-associated protein (CD2AP) serves as a slit diaphragm (SD) protein and plays essential roles in maintaining podocyte integrity and reducing proteinuria. MicroRNAs (miRNAs) are novel regulators of gene expression. Podocyte-specific loss of miRNAs would lead to significant proteinuria. Here, we report new evidence in which miRNAs may function to suppress CD2AP expression through a transcriptional way. By scanning human CD2AP promoter in silico for sequences complementary to known miRNAs, we chose miR-939, miR-148b*, miR-191*, miR-638 as four candidates and transfected them into HEK-293T cells. Dual-luciferase reporter assay identified that only miR-939 significantly reduced the relative luciferase activity of CD2AP promoter region. Further analysis confirmed that the mRNA and protein expressions of CD2AP were also down-regulated by miR-939. In conclusion, we have identified that miR-939 targets CD2AP promoter sequences and suppresses its gene expression. These findings suggest that miRNAs may mediate podocyte injury via reducing the expression of the SD proteins, such as CD2AP.
authors.affiliation: a Department of Pediatrics , the First Affiliated Hospital, Nanjing Medical University , Nanjing , Jiangsu , China ;
authors.forename: Yu-Ping
authors.initials: YP
authors.lastname: Huang
authors.affiliation: b Department of Rheumatology , Nanjing Children's Hospital Affiliated to Nanjing Medical University , Nanjing , Jiangsu , China.
authors.forename: Ling-Zhi
authors.initials: LZ
authors.lastname: Qiu
authors.affiliation: a Department of Pediatrics , the First Affiliated Hospital, Nanjing Medical University , Nanjing , Jiangsu , China ;
authors.forename: Guo-Ping
authors.initials: GP
authors.lastname: Zhou
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000096:biosynthesis*; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000096:biosynthesis*; D015536:Down-Regulation; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000502:physiology*; D050199:Podocytes / Q000502:physiology; D011401:Promoter Regions, Genetic
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C114545:CD2-associated protein; D003598:Cytoskeletal Proteins; C000604729:MIRN939 microRNA, human; D035683:MicroRNAs
keywords: CD2-associated protein; gene expression regulation; microRNA-939; podocyte injury; promoter