title: miR-365, a novel negative regulator of interleukin-6 gene expression, is cooperatively regulated by Sp1 and NF-kappaB.
abstract: Interleukin-6 (IL-6) is a pleiotropic cytokine that plays a central role in host defense. IL-6 expression can be regulated at both a transcriptional and a post-transcriptional level. We used a combination of bioinformatics and experimental techniques to demonstrate that the miR-365 is a direct negative regulator of IL-6. Overexpression of miR-365 mimics decreased activity of a luciferase reporter containing the IL-6 3'-UTR and led to repression of IL-6 protein. In contrast, ectopic expression of a miR-365 inhibitor elevated IL-6 expression. The negative regulation of miR-365 was strictly dependent on a microRNA binding element in the 3'-UTR of IL-6 mRNA. Deletion mutant analysis of the miR-365 promoter showed that two transcription factors, Sp1 and NF-κB, are essential for the transcriptional regulation of miR-365. We also demonstrate that the MAPK/ERK pathway contributes to the regulation of miR-365. Furthermore, miR-365 exhibited a greater negative regulatory effect on IL-6 than hsa-let-7a, a previously identified microRNA negatively regulating IL-6. Taken together, our results show that miR-365 is a novel negative regulator of IL-6.
authors.affiliation: Division of Animal Infectious Diseases, State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Xu
authors.forename: Shao-Bo
authors.initials: SB
authors.lastname: Xiao
authors.forename: Peng
authors.initials: P
authors.lastname: Xu
authors.forename: Qian
authors.initials: Q
authors.lastname: Xie
authors.forename: Lu
authors.initials: L
authors.lastname: Cao
authors.forename: Dang
authors.initials: D
authors.lastname: Wang
authors.forename: Rui
authors.initials: R
authors.lastname: Luo
authors.forename: Yao
authors.initials: Y
authors.lastname: Zhong
authors.forename: Huan-Chun
authors.initials: HC
authors.lastname: Chen
authors.forename: Liu-Rong
authors.initials: LR
authors.lastname: Fang
mesh_terms: D020413:3' Untranslated Regions; D001483:Base Sequence; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis*; D035683:MicroRNAs / Q000096:biosynthesis* / Q000502:physiology*; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D016329:Sp1 Transcription Factor / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D015850:Interleukin-6; C546819:MIRN365 microRNA, human; D035683:MicroRNAs; D016328:NF-kappa B; D034741:RNA, Small Interfering; D016329:Sp1 Transcription Factor