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Role of Ca2+/calmodulin-dependent kinase II-IRAK1 interaction in LMP1-induced NF-κB activation

Jung-Eun Kim1, Sang Yong Kim, Sue Yeon Lim

  • 1Department of Life Science, Gachon University, Seongnam-Si, Kyeonggi-Do, South Korea.

Molecular and Cellular Biology
|November 20, 2013
PubMed

Insights

Interleukin-1 receptor-associated kinase 1 (IRAK1) acts as a scaffold, recruiting Ca2+/calmodulin-dependent protein kinase II (CaMKII) to phosphorylate p65/RelA. This interaction is crucial for Epstein-Barr virus LMP1-induced NF-κB activation.

Area of Science:

  • Molecular Biology
  • Virology
  • Immunology

Background:

  • Interleukin-1 receptor-associated kinase 1 (IRAK1) is essential for Epstein-Barr virus (EBV) latent infection membrane protein 1 (LMP1)-induced NF-κB activation.
  • IRAK1's kinase activity is not required, suggesting a scaffold function for recruiting kinases that phosphorylate p65/RelA at serine 536.
  • This phosphorylation enhances NF-κB-dependent transcriptional activity.

Purpose of the Study:

  • To identify the specific kinase responsible for p65/RelA serine 536 phosphorylation in the context of EBV LMP1 signaling.
  • To elucidate the role of IRAK1 in recruiting and activating this kinase.
  • To understand the mechanism by which IRAK1 and the identified kinase contribute to NF-κB activation.

Main Methods:

  • Investigated the interaction between Ca2+/calmodulin-dependent protein kinase II (CaMKII) and IRAK1.
  • Assessed CaMKII's ability to phosphorylate p65/RelA directly in vitro.
  • Utilized siRNA or small molecule inhibitors to downregulate CaMKII activity/expression and IRAK1 knockout mouse embryonic fibroblasts (MEFs) to evaluate the impact on LMP1-induced signaling.

Main Results:

  • CaMKII was found to interact with the death domain of IRAK1.
  • CaMKII directly phosphorylated p65/RelA at serine 536 in vitro.
  • Downregulation of CaMKII or IRAK1 significantly reduced LMP1-induced p65/RelA serine 536 phosphorylation and NF-κB activation, with reduced CaMKII activation observed in IRAK1 KO MEFs.

Conclusions:

  • IRAK1 functions as a scaffold protein that recruits and activates CaMKII.
  • Activated CaMKII phosphorylates p65/RelA at serine 536, enhancing the transactivation potential of NF-κB.
  • This IRAK1-CaMKII-p65/RelA axis is critical for Epstein-Barr virus LMP1-induced NF-κB activation pathway.

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