MicroRNA-27 enhances differentiation of myeloblasts into granulocytes by post-transcriptionally downregulating Runx1.
We investigated the regulation of the transcription factor Runx1 by microRNA (miR)-27 and the resulting effects upon the differentiation of myeloblasts into granulocytes. When 32D.cl3 cell differentiation was induced using granulocyte colony-stimulating factor (CSF3), Runx1 transcription was moderately downregulated, while Runx1 protein levels were completely inhibited, suggesting an involvement of post-transcriptional regulation. Simultaneously, levels of miR-27 and its precursor increased substantially. Reporter assays revealed that miR-27 targets the 3'UTR of the Runx1 transcript. Furthermore, introduction of pre-miR-27 alone into 32D.cl3 cells resulted in downregulation of Runx1 protein, thereby allowing the cell differentiation even in the absence of CSF3. Conversely, transduction of anti-miR-27 caused upregulation of Runx1 protein, thereby antagonizing the CSF3-mediated granulocyte differentiation. Finally, the CSF3-induced transcription factor C/EBPalpha enhanced transcription of a host gene of miR-27, C9orf3, via activation of its promoter. Thus, miR-27 enhances differentiation of myeloblasts into granulocytes via post-transcriptional downregulation of Runx1.
Feng
Jue
J
Department of Molecular Immunology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan.
Iwama
Atsushi
A
Satake
Masanobu
M
Kohu
Kazuyoshi
K
D020413:3' Untranslated Regions; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D050676:Core Binding Factor Alpha 2 Subunit / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D018274:Electroporation; D015870:Gene Expression; D005786:Gene Expression Regulation; D042381:Granulocyte Precursor Cells / Q000502:physiology*; D006098:Granulocytes / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D049409:Luciferases, Firefly / Q000235:genetics; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology; D016184:Receptors, Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods
D020413:3' Untranslated Regions; C517047:CSF3R protein, human; D050676:Core Binding Factor Alpha 2 Subunit; C531388:MIRN27 microRNA, human; D035683:MicroRNAs; D034741:RNA, Small Interfering; D016184:Receptors, Colony-Stimulating Factor; D049409:Luciferases, Firefly