ZNF216 Is an A20-like and IkappaB kinase gamma-interacting inhibitor of NFkappaB activation

J Biol Chem. 2004 Apr 16;279(16):16847-53. doi: 10.1074/jbc.M309491200. Epub 2004 Jan 30.


The transcription factor NFkappaB plays important roles in immune regulation, inflammatory responses, and anti-apoptosis. Activation of NFkappaB requires the activity of IkappaB kinase, a kinase complex that contains two catalytic subunits, IKKalpha and IKKbeta, and a non-enzymatic regulatory subunit, IKKgamma. To understand how NFkappaB activation is regulated at the IKKgamma level, we searched for IKKgamma-interacting proteins by the yeast two-hybrid system. This search identified ZNF216, a zinc finger protein with unknown biological functions. ZNF216 contains an A20-like zinc finger domain (ZnF-A20) at its N terminus and an AN1-like domain (ZnF-AN1) at its C terminus. Similar to A20, ZNF216 interacted with IKKgamma, RIP, and TRAF6 in co-immunoprecipitation experiments. Domain mapping experiments indicated that the ZnF-A20 domain was responsible for interacting with IKKgamma and RIP, whereas the ZnF-AN1 domain interacted with TRAF6. ZNF216 inhibited NFkappaB activation triggered by overexpression of RIP and TRAF6 but not of p65. ZNF216 also inhibited tumor necrosis factor (TNF)-, interleukin-1-, and Toll-like receptor 4-induced NFkappaB activation in a dose-dependent manner. The ZnF-A20 domain was essential for ZNF216-mediated inhibition of NFkappaB activation. The ZnF-A20 and ZnF-AN1 domains of ZNF216 could interact with each other, whereas ZNF216 could form homo-oligomers or hetero-oligomers with A20. Unlike A20, which inhibits TNF-induced apoptosis, overexpression of ZNF216 sensitized cells to TNF-induced apoptosis. Our findings suggest that ZNF216 and A20 have redundant and distinct roles in regulating NFkappaB activation and apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Apoptosis
  • DNA-Binding Proteins
  • Humans
  • I-kappa B Kinase
  • Intracellular Signaling Peptides and Proteins
  • NF-kappa B / metabolism*
  • Nuclear Proteins
  • Phosphorylation
  • Protein Binding
  • Protein Serine-Threonine Kinases / antagonists & inhibitors*
  • Protein Serine-Threonine Kinases / metabolism
  • Proteins / genetics
  • Proteins / metabolism*
  • Signal Transduction
  • Tumor Necrosis Factor alpha-Induced Protein 3


  • DNA-Binding Proteins
  • Intracellular Signaling Peptides and Proteins
  • NF-kappa B
  • Nuclear Proteins
  • Proteins
  • ZFAND5 protein, human
  • Protein Serine-Threonine Kinases
  • CHUK protein, human
  • I-kappa B Kinase
  • IKBKB protein, human
  • IKBKE protein, human
  • TNFAIP3 protein, human
  • Tumor Necrosis Factor alpha-Induced Protein 3