Isolation of the chicken NF-kappa B p65 subunit-encoding cDNA and characterization of its products.
NF-kappa B is a heterodimeric transcription factor consisting of subunits of 50 kDa (p50) and 65 kDa (p65). cDNA clones encoding the chicken NF-kappa B p65 subunit were isolated. Sequence analysis showed that chicken p65 is approximately 55% identical to the mouse and human p65 proteins, and contains the Rel homology domain (RHD) in its N-terminal 286 amino acids (aa) and the putative transactivation domain in its C-terminal region. The RHD is particularly highly conserved between the chicken and mammalian p65 proteins. Northern blot hybridization analysis detected the expression of a 2.6-kb transcript of p65 in various organs, with the highest level in spleen. A fusion protein containing the RHD of chicken p65 was found to bind to a consensus kappa B-site in an electrophoretic mobility shift assay (EMSA). This binding was specifically inhibited by the presence of fusion proteins containing the C-terminal ankyrin repeats domain (ARD) of chicken p105, the precursor protein for the p50 subunit. Immunoprecipitation analysis showed that p65 formed a complex(es) with multiple cellular proteins, including p50, p105 and c-Rel in chicken spleen cells.
Ikeda
T
T
National Institute of Animal Health, Ibaraki, Japan.
Honjo
K
K
Hirota
Y
Y
Onodera
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002645:Chickens; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D004926:Escherichia coli; D006801:Humans; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D017386:Sequence Homology, Amino Acid; D051996:Transcription Factor RelA
D016328:NF-kappa B; D051996:Transcription Factor RelA; D004247:DNA