Allele-31 C>T regulates binding activity to IL-1βgene promoter of nuclear transcription factor C/EBPβand PU.1 induced by Mycobacterium tuberculosis infection
10.3760/cma.j.issn.1674-2397.2016.02.016
- VernacularTitle:结核分枝杆菌诱导的核转录因子C／EBPβ和PU．1通过等位基因-31C＞T调控IL-1β基因启动子活性
- Author:
Guoliang ZHANG
;
Rongrong ZOU
;
Lingling WANG
;
Wenfei WANG
;
Mingxia ZHANG
;
Guilin YANG
;
Xinchun CHEN
- Publication Type:Journal Article
- Keywords:
Interleukin-1beta;
Alleles;
Nuclear transcription factors;
Promoter;
Mycobacterium tuberculosis
- From:Chinese Journal of Clinical Infectious Diseases
2016;9(2):180-185
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of allele-31 C>T on the binding activity to IL-1βpromoter of the nuclear transcription factor C/EBPβand PU.1 induced by Mycobacterium tuberculosis infection.Methods The electrophoretic mobility shift assay ( EMSA) was performed to explore whether the nuclear transcription factor C/EBPβand PU.1 could bind to -31 region in IL-1βpromoter.The C/EBPβ-and PU.1-expressing vectors were constructed and co-transfected into HeLa cells with IL-1βpromoter luciferase vector.The expression of C/EBPβand PU.1 was confirmed using Western blotting assay, and the promoter activity was determined using Dual-Glo Luciferase system under various transfection conditions. Lentivirus-mediated RNA interference was used to explore the effects of C/EBPβand PU.1 on IL-1βexpression.GraphPad Prism 5.0 was used for data analysis.Results EMSA results showed that both C/EBPβand PU.1 could bind to -31 region in IL-1βpromoter.Both C/EBPβand PU.1 induced by Mycobacterium tuberculosis infection could increase IL-1βpromoter activity, especially for the -31 T allele (t=22.33 and 7.98,P<0.01), and there was a synergy on the promoter activity between C/EBPβand PU.1.The promoter activity decreased significantly when C/EBPβand/or PU.1 were silenced by lentivirus-mediated RNA interference (q=5.79, 6.23 and 11.66,P<0.01).Conclusion The allele-31 C>T can induce IL-1βpromoter activity and gene transcription through regulation of binding activity to C/EBPβand PU.1 induced by Mycobacterium tuberculosis infection.
