Open AccessInhibition of Canonical NF-κB Nuclear Localization by (−)-DHMEQ via Impairment of DNA Binding
Author(s) -
Kana Horie,
Jun Ma,
Kazuo Umezawa
Publication year - 2015
Publication title -
oncology research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.267
H-Index - 57
eISSN - 1555-3906
pISSN - 0965-0407
DOI - 10.3727/096504014x14146137738628
Subject(s) - relb , nuclear localization sequence , nuclear protein , cell nucleus , importin , microbiology and biotechnology , dna , biology , mutant , nf κb , chemistry , cytoplasm , nfkb1 , nuclear transport , biochemistry , transcription factor , signal transduction , gene
We previously discovered (-)-DHMEQ as a selective inhibitor of NF-κB, and it was shown to suppress many cancer and inflammation models in animals. (-)-DHMEQ directly binds to NF-κB components to inhibit DNA binding, and moreover, it often inhibits nuclear translocation of NF-κB. The mechanism of inhibiting nuclear translocation has been elucidated for RelB, a main noncanonical NF-κB component. However, it was not elucidated for p65, a main canonical NF-κB component. In the present research, we studied how (-)-DHMEQ inhibits nuclear localization of p65. First, (-)-DHMEQ inhibited p65 nuclear accumulation in adult T-cell leukemia MT-2 cells in which canonical p65 is constitutively activated. But there was no change in the stability and importin-α3 affinity of p65. Then, we prepared a p65 mutant protein with Arg35Ala and Tyr36Ala (AA) mutations having no DNA-binding ability in HeLa cells. The p65 AA mutant showed reduced nuclear localization without changing the stability and importin affinity. Taken together, the mechanism of inhibition is different between RelB and p65, and inhibition of p65 nuclear localization is likely to be due to the inhibition of DNA binding changing the equilibrium between the nuclear and cytoplasmic amounts of p65.