Essential role of C/EBPalpha in G-CSF-induced transcriptional activation and chromatin modification of myeloid-specific genes.
Granulocyte colony-stimulating factor (G-CSF) regulates the proliferation and differentiation of neutrophilic progenitor cells. Here, we investigated the roles of CCAAT/enhancer-binding protein (C/EBP)alpha in the G-CSF-induced transcriptional activation and chromatin modification of the CCR2 and myeloperoxidase (MPO) genes in IL-3-dependent myeloid FDN1.1 cells. Chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assays revealed that G-CSF activates C/EBPalpha to bind target promoters. ChIP mapping experiments across the CCR2 and MPO genes showed that G-CSF induces histone H3 modifications: the acetylation of Lys9, trimethylation of Lys4 and trimethylation of Lys9. The distribution profile of the trimethylated Lys9 was distinct from that of the two other modifications. All the G-CSF-induced C/EBPalpha recruitment, transcriptional activation and histone modifications were reversed by re-stimulation with IL-3, and were abolished by short hairpin RNA (shRNA)-mediated knockdown of C/EBPalpha. These results indicate that C/EBPalpha is activated by G-CSF to bind target promoters, and plays critical roles in the transcriptional activation and dynamic chromatin modification of target genes during neutrophil differentiation.
Iida
Satoshi
S
Department of Medical Chemistry, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Watanabe-Fukunaga
Rie
R
Nagata
Shigekazu
S
Fukunaga
Rikiro
R
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000378:metabolism*; D002460:Cell Line; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects; D047369:Chromatin Immunoprecipitation; D017931:DNA Primers / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D006657:Histones / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D038042:Myelopoiesis / Q000187:drug effects* / Q000235:genetics* / Q000502:physiology; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009195:Peroxidase / Q000235:genetics; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics; D054390:Receptors, CCR2 / Q000235:genetics; D011994:Recombinant Proteins; D015533:Transcriptional Activation / Q000187:drug effects
D022763:CCAAT-Enhancer-Binding Protein-alpha; C515335:Ccr2 protein, mouse; D017931:DNA Primers; D006657:Histones; D034741:RNA, Small Interfering; D054390:Receptors, CCR2; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D009195:Peroxidase