NF kappa B- and AP-1-mediated DNA looping regulates matrix metalloproteinase-9 transcription in TNF-alpha-treated human leukemia U937 cells

作者：Chen, Ying Jung; Chang, Long Sen^*

来源：Biochimica et Biophysica Acta-Gene Regulatory Mechanisms, 2015, 1849(10): 1248-1259.

DOI：10.1016/j.bbagrm.2015.07.016

• MMP-9
• DNA looping
• TNF-alpha
• NF kappa B/p65
• AP-1/c-Jun
• Histone acetylation

The aim of this study is to explore the spatial association of critical genomic elements in the effect of TNF-alpha on matrix metalloproteinase-9 (MMP-9) expression in human leukemia U937 cells. TNF-alpha, up-regulated MMP-9 protein expression and mRNA level in U937 cells, and Akt-mediated-NF kappa B/p65 activation and JNK-mediated c-Jun activation were proven to be involved in TNF-alpha-induced MMP-9 up-regulation. Promoter luciferase activity assay revealed that NF kappa B (nt-600) and AP-1 (nt-79) binding sites were crucial for TNF-alpha-induced transcription of MMP-9 gene. The results of a chromatin immunoprecipitation assay indicated that TNF-alpha reduced histone deacetylase-1 (HDAC-1) recruitment but increased p300 (a histone acetyltransferase) recruitment to MMP-9 promoter regions surrounding NF kappa B and AP-1 binding sites. Consistently, TNF-alpha increased enrichment of the acetylated histone H3 mark on MMP-9 promoter regions. DNA affinity purification assay revealed that p300 and HDAC1 could bind oligonucleotides containing AP-1/c-Jun and NF kappa B/p65 binding sites. Chromosome conformation capture assay showed that TNF-alpha stimulated chromosomal loops in the MMP-9 promoter via NF kappa B/p65 and AP-1/c-Jun. The p300-associated acetyltransferase activity was crucial for p65/c-Jun-mediated DNA looping, and inhibition of HDAC activity increased the level of DNA looping. Reduction in the level of DNA looping eliminated all TNF-alpha-stimulated MMP-9 up-regulation. Taken together, our data suggest that p65/c-Jun-mediated DNA looping is involved in TNF-alpha-induced MMP-9 up-regulation and that the recruitment of p300 or HDAC1 to NF kappa B and AP-1 binding sites modifies the level of DNA looping.

• 出版日期2015-10
• 单位中山大学