Inhibition of nuclear factor-kappaB-mediated transcription by association with the amino-terminal enhancer of split, a Groucho-related protein lacking WD40 repeats.
The amino-terminal enhancer of split (AES) encodes a 197-amino acid protein that is homologous to the NH(2)-terminal domain of the Drosophila Groucho protein but lacks COOH-terminal WD40 repeats. Although the Drosophila Groucho protein and its mammalian homologs, transducin-like enhancer of split proteins, are known to act as non-DNA binding corepressors, the role of the AES protein remains unclarified. Using the yeast two-hybrid system, we have identified the protein-protein interaction between AES and the p65 (RelA) subunit of the transcription factor nuclear factor kappaB (NF-kappaB), which activates various target genes involved in inflammation, apoptosis, and embryonic development. The interaction between AES and p65 was confirmed by in vitro glutathione S-transferase pull down assay and by in vivo co-immunoprecipitation study. In transient transfection assays, AES repressed p65-driven gene expression. AES also inhibited NF-kappaB-dependent gene expression induced by tumor necrosis factor alpha, interleukin-1beta, and mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase 1, which is an upstream kinase for NF-kappaB activation. These data indicate that AES acts as a corepressor for NF-kappaB and suggest that AES may play a pivotal role in the regulation of NF-kappaB target genes.
Tetsuka
T
T
Department of Molecular Genetics, Nagoya City University Medical School, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, Aichi 467-8601, Japan.
Uranishi
H
H
Imai
H
H
Ono
T
T
Sonta
S
S
Takahashi
N
N
Asamitsu
K
K
Okamoto
T
T
D051792:Basic Helix-Loop-Helix Proteins; D056970:Co-Repressor Proteins; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D005982:Glutathione Transferase / Q000235:genetics; D015497:HIV-1 / Q000235:genetics; D015801:Helminth Proteins / Q000235:genetics*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D015850:Interleukin-6 / Q000235:genetics; D019169:Jurkat Cells; D048728:MAP Kinase Kinase Kinase 1; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011233:Precipitin Tests; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism* / Q000494:pharmacology; D012097:Repressor Proteins / Q000378:metabolism*; D051996:Transcription Factor RelA; D014158:Transcription, Genetic; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D015003:Yeasts
D051792:Basic Helix-Loop-Helix Proteins; D056970:Co-Repressor Proteins; D004268:DNA-Binding Proteins; D005982:Glutathione Transferase; D015801:Helminth Proteins; D007375:Interleukin-1; D015850:Interleukin-6; D048728:MAP Kinase Kinase Kinase 1; D016328:NF-kappa B; D017346:Protein Serine-Threonine Kinases; D011506:Proteins; D012097:Repressor Proteins; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; C498445:TLE5 protein, human; C400683:che-2 protein, C elegans; C091023:gro protein, Drosophila; C482576:MAP3K1 protein, human