Isolation of a rel-related human cDNA that potentially encodes the 65-kD subunit of NF-kappa B

阅读量：

作者：

S Ruben，P Dillon，R Schreck，T Henkel，C Chen，M Maher，P Baeuerle，C Rosen

展开

摘要：

A DNA probe that spanned a domain conserved among the proto-oncogene c-rel, the Drosophila morphogen dorsal, and the p50 DNA binding subunit of NF-κB was generated from Jurkat T cell complementary DNA with the polymerase chain reaction (PCR) and degenerate oligonucleotides. This probe was used to identify a rel-related complementary DNA that hybridized to a 2.6-kilobase messenger RNA present in human T and B lymphocytes. In vitro transcription and translation of the complementary DNA resulted in the synthesis of a protein with an apparent molecular size of 65 kilodaltons (kD). The translated protein showed weak DNA binding with a specificity for the κB binding motif. This protein-DNA complex comigrated with the complex obtained with the purified human p65 NF-κB subunit and binding was inhibited by IκB-α and -β proteins. In addition, the 65-kD protein associated with the p50 subunit of NF-κ B and the κB probe to form a complex with the same electrophoretic mobility as the NF-κ B-DNA complex. Therefore the rel-related 65-kD protein may represent the p65 subunit of the active NF-κB transcription factor complex.

展开

关键词：

T-Lymphocytes Humans Macromolecular Substances NF-kappa B Proto-Oncogene Proteins c-rel Proto-Oncogene Proteins DNA Blotting, Northern Cloning, Molecular Polymerase Chain Reaction