Premium
In vitro transcription assay with the purified 40kDa NF1‐like protein binding to the rat p53 promoter
Author(s) -
Lee Minhyung,
Song Haisun,
Lee Kyunghee,
Park Jongsang
Publication year - 1999
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549900201453
Subject(s) - microbiology and biotechnology , ap 1 transcription factor , promoter , transactivation , transcription (linguistics) , transcription factor , dna footprinting , taf2 , in vitro , general transcription factor , biology , e box , activator (genetics) , chemistry , dna binding protein , gene , biochemistry , gene expression , linguistics , philosophy
The rat p53 promoter has several potential transcription factor‐recognition motifs. They inculde NF1‐like, bHLH family, and AP1‐like proteins binding sites. The binding protein to NF1‐like motif was previously identified. The protein has about 40kDa of molecular mass, which is smaller than that of NF1. Anti‐NF1 polyclonal antibody does not recognize the protein. In this study, we isolated the 40kDa protein by sequence‐specific DNA affinity chromatography. The isloated protein was assayed by DNase I footprinting analysis. To determine the transactivation effect of the protein, in vitro transcription with the purified 40kDa protein was carried out. After the addition of the purified 40kDa protein into the transcription reaction mixture, the transcription level of the p53 promoter was increased. This suggests that the 40 kDa NF1‐like protein is a transcription activator for the rat p53 gene.