Purification of the major histocompatibility complex class I transcription factor H2TF1. The full-length product of the nfkb2 gene.
H2TF1 is a ubiquitous major histocompatibility complex (MHC) class I-specific transcription factor, which binds to the palindromic kappa B enhancer site upstream of MHC class I genes. Here we report that H2TF1 consists of a polypeptide with relative molecular mass 110,000, that corresponds to the predicted 100-kDa product (NF-kappa B2 p100) encoded by the candidate proto-oncogene nfkb2 (lyt-10). H2TF1 was purified by a novel affinity chromatography method and identified as the NF-kappa B2 p100 polypeptide by peptide sequencing as well as by reactivity with a specific antiserum. Purified H2TF1 binds the MHC kappa B site with high affinity (KD = 3 x 10(-11) M), in contrast with previous reports that NF-kappa B2 p100 did not bind DNA.
Potter
D A
DA
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.
Larson
C J
CJ
Eckes
P
P
Schmid
R M
RM
Nabel
G J
GJ
Verdine
G L
GL
Sharp
P A
PA
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002846:Chromatography, Affinity; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005805:Genes, MHC Class I; D006367:HeLa Cells / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016328:NF-kappa B; D052003:NF-kappa B p52 Subunit; D010777:Photochemistry; D000090063:Proto-Oncogene Mas; D017421:Sequence Analysis; D014157:Transcription Factors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D014466:Ultraviolet Rays
C000715470:MAS1 protein, human; D016328:NF-kappa B; D052003:NF-kappa B p52 Subunit; D000090063:Proto-Oncogene Mas; D014157:Transcription Factors; D004247:DNA