Identification of an NF-Y/HMG-I(Y)-binding site in the human IL-10 promoter.
The cis-acting elements and transcriptional factors that control interleukin 10 (IL-10) promoter activity remain to be clarified. In this report, by performing electrophoresis mobility shift assay (EMSA) with nuclear extracts from the Raji B cell line, the nuclear factor Y (NF-Y) was found to be capable of binding to a CCAAT box-containing region between the -215 and -192bps (the -215/-192 region), relative to the transcription initiation site, in the human IL-10 promoter. The binding of NF-Y to the promoter appears to be enhanced by the presence of HMG-I(Y), since anti-HMG-I(Y) antibody diminished the NF-Y-binding activity in EMSA. Two T nucleotides at -192 and -193bps were found to be critical for the binding of NF-Y and HMG-I(Y) to the -215/-192 promoter region, suggesting the possibility that HMG-I(Y) binds to the AT-rich promoter region downstream of the CCAAT box at the -210/-199 region. These findings suggest that NF-Y and HMG-I(Y) may play an important role in regulating the IL-10 promoter activity in B cells.
Lin
Shih-Chang
SC
Division of Allergy and Immunology, Department of Internal Medicine, Cathay General Hospital, Taipei, Taiwan. sclin@cgh.org.tw
D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D023081:CCAAT-Binding Factor / Q000378:metabolism*; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism; D025741:HMGA1a Protein / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D016753:Interleukin-10 / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding
D023081:CCAAT-Binding Factor; D025741:HMGA1a Protein; D016753:Interleukin-10; D004247:DNA