title: A dual cis-regulatory code links IRF8 to constitutive and inducible gene expression in macrophages.
abstract: The transcription factor (TF) interferon regulatory factor 8 (IRF8) controls both developmental and inflammatory stimulus-inducible genes in macrophages, but the mechanisms underlying these two different functions are largely unknown. One possibility is that these different roles are linked to the ability of IRF8 to bind alternative DNA sequences. We found that IRF8 is recruited to distinct sets of DNA consensus sequences before and after lipopolysaccharide (LPS) stimulation. In resting cells, IRF8 was mainly bound to composite sites together with the master regulator of myeloid development PU.1. Basal IRF8-PU.1 binding maintained the expression of a broad panel of genes essential for macrophage functions (such as microbial recognition and response to purines) and contributed to basal expression of many LPS-inducible genes. After LPS stimulation, increased expression of IRF8, other IRFs, and AP-1 family TFs enabled IRF8 binding to thousands of additional regions containing low-affinity multimerized IRF sites and composite IRF-AP-1 sites, which were not premarked by PU.1 and did not contribute to the basal IRF8 cistrome. While constitutively expressed IRF8-dependent genes contained only sites mediating basal IRF8/PU.1 recruitment, inducible IRF8-dependent genes contained variable combinations of constitutive and inducible sites. Overall, these data show at the genome scale how the same TF can be linked to constitutive and inducible gene regulation via distinct combinations of alternative DNA-binding sites.
authors: Department of Experimental Oncology, European Institute of Oncology (IEO), 20139 Milan, Italy;
authors: Alessandra
authors: A
authors: Mancino
authors: Department of Experimental Oncology, European Institute of Oncology (IEO), 20139 Milan, Italy;
authors: Alberto
authors: A
authors: Termanini
authors: Department of Experimental Oncology, European Institute of Oncology (IEO), 20139 Milan, Italy;
authors: Iros
authors: I
authors: Barozzi
authors: Department of Experimental Oncology, European Institute of Oncology (IEO), 20139 Milan, Italy;
authors: Serena
authors: S
authors: Ghisletti
authors: Department of Experimental Oncology, European Institute of Oncology (IEO), 20139 Milan, Italy;
authors: Renato
authors: R
authors: Ostuni
authors: Department of Experimental Oncology, European Institute of Oncology (IEO), 20139 Milan, Italy;
authors: Elena
authors: E
authors: Prosperini
authors: Laboratory of Molecular Growth Regulation, Genomics of Differentiation Program, National Institute of Child Health and Human Development (NICHD), National Institutes of Health, Bethesda, Maryland 20892, USA.
authors: Keiko
authors: K
authors: Ozato
authors: Department of Experimental Oncology, European Institute of Oncology (IEO), 20139 Milan, Italy; gioacchino.natoli@ieo.eu.
authors: Gioacchino
authors: G
authors: Natoli
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D050835:Interferon Regulatory Factors / Q000235:genetics / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D011485:Protein Binding; D000099185:Interferon Regulatory Factor-8
chemical_list: D050835:Interferon Regulatory Factors; D008070:Lipopolysaccharides; D000099185:Interferon Regulatory Factor-8
keywords: IRF8; chromatin; epigenome; inflammation; interferon; macrophages; transcription
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