NF-κB- and AP-1-mediated DNA looping regulates osteopontin transcription in endotoxin-stimulated murine macrophages.
Osteopontin (OPN) is expressed by various immune cells and modulates both innate and adaptive immune responses. However, the molecular mechanisms that control opn gene expression, especially at the chromatin level, remain largely unknown. We have previously demonstrated many specific cis- and trans-regulatory elements that determine the extent of endotoxin (LPS)-mediated induction of OPN synthesis in murine macrophages. In the present study, we confirm that NF-κB also plays an important role in the setting of LPS-stimulated OPN expression through binding to a distal regulatory element. Importantly, we demonstrate that LPS stimulates chromosomal loops in the OPN promoter between NF-κB binding site and AP-1 binding site using chromosome conformation capture technology. The crucial role of NF-κB and AP-1 in LPS-stimulated DNA looping was confirmed, as small interfering RNA knock-down of NF-κB p65 and AP-1 c-Jun exhibited decreased levels of DNA looping. Furthermore, we demonstrate that p300 can form a complex with NF-κB and AP-1 and is involved in DNA looping and LPS-induced OPN expression. Therefore, we have identified an essential mechanism to remodel the local chromatin structures and spatial conformations to regulate LPS-induced OPN expression.
Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University Medical School, Jinan, Shandong 250012, China.
Wei
W
Zhao
Lijuan
L
Wang
Meng
M
Zhang
Peng
P
Wang
Lei
L
Zhang
Chao
C
Yuan
Jianni
J
Qi
Yu
Y
Qiao
Paul C
PC
Kuo
Chengjiang
C
Gao
D000818:Animals; D002460:Cell Line; D004247:DNA / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000502:physiology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000096:biosynthesis; D005786:Gene Expression Regulation / Q000276:immunology; D051550:I-kappa B Kinase / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000502:physiology*; D008262:Macrophage Activation / Q000235:genetics* / Q000276:immunology; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D009690:Nucleic Acid Conformation; D053495:Osteopontin / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D051996:Transcription Factor RelA / Q000037:antagonists & inhibitors; D014158:Transcription, Genetic / Q000276:immunology*; D050880:p300-CBP Transcription Factors / Q000378:metabolism
D008070:Lipopolysaccharides; D016328:NF-kappa B; D016755:Proto-Oncogene Proteins c-jun; D034741:RNA, Small Interfering; C498144:Rela protein, mouse; D018808:Transcription Factor AP-1; D051996:Transcription Factor RelA; D053495:Osteopontin; D004247:DNA; D050880:p300-CBP Transcription Factors; D051550:I-kappa B Kinase; D048049:Extracellular Signal-Regulated MAP Kinases
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