[Brd3 promotes IL-6 production via enhancing acetylase CBP recruitment and histone 3 acetylation within IL6 promoter].
Objective To investigate the role of bromodomain containing 3 (Brd3) in LPS-triggered interleukin-6 (IL-6) production in macrophages and the underlying mechanism. Methods CRISPR-Cas9 technology was used to screen an RAW264.7 cell line with Brd3 knockout (Brd3-/-). The Brd3-/-cells were used as an experimental group, and the parential cells expressing wide-type Brd3 as a control group. The IL-6 level in cell culture supernatant was detected by ELISA after 100 ng/mL LPS challenging. Effect of Brd3 knockout on the expression and activation of signal pathways involved in IL-6 expression, including the NF-κB and mitogen-activated protein kinase (MAPK) pathways were examined by Western blot analysis. Chromatin immunoprecipitation (ChIP) assay was used to evaluate the recruitment of acetylase CREB-binding protein (CBP) to IL6 gene promoter and the acetylation level of histone 3 within IL6 gene promoter. Results LPS treatment significantly downregulated Brd3 expression in mouse peritoneal macrophages. LPS-induced production of IL-6 was significantly inhibited in Brd3-/-macrophages. The expressions and activation of signal molecules within NF-κB and MAPK pathways were barely affected. Brd3 knockout significantly decreased the recruitment of acetylase CBP to IL6 gene promoter, and the acetylation level of histone3 within IL6 gene promoter was also repressed. Conclusion Brd3 promotes LPS-triggered IL-6 production via promoting the recruitment of CBP to IL6 promoter and enhancing the acetylation level of histone 3 within IL6 promoter.
Department of Immunology, College of Basic Medicine, Second Military Medical University, Shanghai 200433, China.
Wenhui
W
Ren
Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Beijing 100005, China.
Donghao
D
Sun
Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Beijing 100005, China. *Corresponding authors, E-mail: wangcm1977@126.com.
Chunmei
C
Wang
Department of Immunology, College of Basic Medicine, Second Military Medical University, Shanghai 200433, China . *Corresponding authors, linan@immunol.org.
Nan
N
Li
D000107:Acetylation; D000818:Animals; D050882:CREB-Binding Protein / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D014157:Transcription Factors; D000092468:Bromodomain Containing Proteins
D050882:CREB-Binding Protein; D006657:Histones; D015850:Interleukin-6; D009687:Nuclear Proteins; D014157:Transcription Factors; C518476:Brd3 protein, mouse; D000092468:Bromodomain Containing Proteins