Dimer of arfaptin 2 regulates NF-κB signaling by interacting with IKKβ/NEMO and inhibiting IKKβ kinase activity

Dong Joo You, Cho Rong Park, Michael Furlong, Okjae Koo, Cheolju Lee, Curie Ahn, Jae Young Seong, Jong-Ik Hwang

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

IκB kinases (IKKs) are a therapeutic target due to their crucial roles in various biological processes, including the immune response, the stress response, and tumor development. IKKs integrate various upstream signals that activate NF-κB by phosphorylating IκB and also regulate many proteins related to cell growth and metabolism. Although they function as a heteromeric complex comprised of kinase subunits and an adaptor, these kinases produce distinct cellular responses by phosphorylating different target molecules, suggesting that they may also be regulated in a subtype-specific manner. In this study, arfaptin 2 was identified as an IKKβ-specific binding partner. Interestingly, arfaptin 2 also interacted with NEMO. Domain mapping studies revealed that the C-terminal region, including the IKKβ HLH domain and the first coiled-coil NEMO region were respectively required for interactions with the arfaptin 2 N-terminal flexible region. Overexpression of arfaptin 2 inhibited tumor necrosis factor (TNF)-α-stimulated nuclear factor-κB (NF-κB) signaling, whereas downregulation of arfaptin 2 by small interfering RNA enhanced NF-κB activity. Dimerization of arfaptin 2 through the Bin-Amphiphysin-Rvs domain may be essential to inhibit activation of NF-κB through multimodal interactions with IKKβs or IKKβ/NEMO, as ectopic expression of the arfaptin 2 fragment responsible for IKK interactions did not change TNFα-stimulated NF-κB activation. These data indicate that arfaptin 2 is the first molecule to regulate NF-κB signaling by interacting with the functional IKK complex but not by direct inhibiting IKKβ phosphorylation.

Original languageEnglish
Pages (from-to)2173-2181
Number of pages9
JournalCellular Signalling
Volume27
Issue number11
DOIs
Publication statusPublished - 2015 Nov 1

Fingerprint

Phosphotransferases
Tumor Necrosis Factor-alpha
Biological Phenomena
Dimerization
Small Interfering RNA
Down-Regulation
Phosphorylation
Growth
Neoplasms
Proteins
Therapeutics
Ectopic Gene Expression
amphiphysin

Keywords

  • Arfaptin 2
  • BAR domain
  • IκB kinases
  • NF-κB
  • TNF-α

ASJC Scopus subject areas

  • Cell Biology

Cite this

Dimer of arfaptin 2 regulates NF-κB signaling by interacting with IKKβ/NEMO and inhibiting IKKβ kinase activity. / You, Dong Joo; Park, Cho Rong; Furlong, Michael; Koo, Okjae; Lee, Cheolju; Ahn, Curie; Seong, Jae Young; Hwang, Jong-Ik.

In: Cellular Signalling, Vol. 27, No. 11, 01.11.2015, p. 2173-2181.

Research output: Contribution to journalArticle

You, Dong Joo ; Park, Cho Rong ; Furlong, Michael ; Koo, Okjae ; Lee, Cheolju ; Ahn, Curie ; Seong, Jae Young ; Hwang, Jong-Ik. / Dimer of arfaptin 2 regulates NF-κB signaling by interacting with IKKβ/NEMO and inhibiting IKKβ kinase activity. In: Cellular Signalling. 2015 ; Vol. 27, No. 11. pp. 2173-2181.
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AB - IκB kinases (IKKs) are a therapeutic target due to their crucial roles in various biological processes, including the immune response, the stress response, and tumor development. IKKs integrate various upstream signals that activate NF-κB by phosphorylating IκB and also regulate many proteins related to cell growth and metabolism. Although they function as a heteromeric complex comprised of kinase subunits and an adaptor, these kinases produce distinct cellular responses by phosphorylating different target molecules, suggesting that they may also be regulated in a subtype-specific manner. In this study, arfaptin 2 was identified as an IKKβ-specific binding partner. Interestingly, arfaptin 2 also interacted with NEMO. Domain mapping studies revealed that the C-terminal region, including the IKKβ HLH domain and the first coiled-coil NEMO region were respectively required for interactions with the arfaptin 2 N-terminal flexible region. Overexpression of arfaptin 2 inhibited tumor necrosis factor (TNF)-α-stimulated nuclear factor-κB (NF-κB) signaling, whereas downregulation of arfaptin 2 by small interfering RNA enhanced NF-κB activity. Dimerization of arfaptin 2 through the Bin-Amphiphysin-Rvs domain may be essential to inhibit activation of NF-κB through multimodal interactions with IKKβs or IKKβ/NEMO, as ectopic expression of the arfaptin 2 fragment responsible for IKK interactions did not change TNFα-stimulated NF-κB activation. These data indicate that arfaptin 2 is the first molecule to regulate NF-κB signaling by interacting with the functional IKK complex but not by direct inhibiting IKKβ phosphorylation.

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