Involvement of DNA binding domain in the cellular stability and importin affinity of NF-κB component RelB

Masatoshi Takeiri, Kana Horie, Daisuke Takahashi, Mariko Watanabe, Ryoichi Horie, Siro Simizu, Kazuo Umezawa

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

NF-κB is a transcription factor for the immune activation and tissue stability, but excess activation of NF-κB often causes inflammation and cancer. An NF-κB component RelB is involved in B-cell maturation and autoimmunity. In the present research we studied the role of the RelB DNA binding domain on cellular stability and importin affinity. We prepared a RelB protein mutated at Arg141 to Ala and Tyr142 to Ala (AA mutant) having no DNA binding activity. The stability of this mutant protein was greatly reduced compared with that of the wild-type protein. We also constructed a nuclear localization signal-inactivated mutant of RelB, and found that this mutant was also unstable in the cells. Thus, RelB destabilization was caused by the loss of DNA binding possibly because of the change in cellular localization. The mutation also decreased the affinity to importin-α5 decreasing the nuclear localization. Our newly discovered NF-κB inhibitor (-)-DHMEQ binds to a specific Cys residue in RelB to inhibit DNA binding and also decreased the stability and importin affinity. These findings would indicate that the DNA binding activity of this transcription factor is a crucial for its stability and intracellular localization.

Original languageEnglish
Pages (from-to)3053-3059
Number of pages7
JournalOrganic and Biomolecular Chemistry
Volume10
Issue number15
DOIs
Publication statusPublished - 2012 Apr 21

Fingerprint

Karyopherins
affinity
deoxyribonucleic acid
DNA
proteins
Transcription Factors
Chemical activation
activation
Nuclear Localization Signals
destabilization
Mutant Proteins
mutations
Autoimmunity
inhibitors
Proteins
B-Lymphocytes
cancer
Cells
Tissue
Inflammation

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Biochemistry

Cite this

Involvement of DNA binding domain in the cellular stability and importin affinity of NF-κB component RelB. / Takeiri, Masatoshi; Horie, Kana; Takahashi, Daisuke; Watanabe, Mariko; Horie, Ryoichi; Simizu, Siro; Umezawa, Kazuo.

In: Organic and Biomolecular Chemistry, Vol. 10, No. 15, 21.04.2012, p. 3053-3059.

Research output: Contribution to journalArticle

Takeiri, Masatoshi ; Horie, Kana ; Takahashi, Daisuke ; Watanabe, Mariko ; Horie, Ryoichi ; Simizu, Siro ; Umezawa, Kazuo. / Involvement of DNA binding domain in the cellular stability and importin affinity of NF-κB component RelB. In: Organic and Biomolecular Chemistry. 2012 ; Vol. 10, No. 15. pp. 3053-3059.
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