Analysis of the NF-kappaB p50 dimer interface by diversity screening.
J Mol Biol
; 310(3): 563-75, 2001 Jul 13.
Article
em En
| MEDLINE
| ID: mdl-11439024
ABSTRACT
An in vivo screen has been devised for NF-kappaB p50 activity in Escherichia coli exploiting the ability of the mammalian transcription factor to emulate a prokaryotic repressor. Active intracellular p50 was shown to repress the expression of a green fluorescent protein reporter gene allowing for visual screening of colonies expressing active p50 on agar plates. A library of mutants was constructed in which the residues Y267, L269, A308 and V310 of the dimer interface were simultaneously randomised and twenty-five novel functional interfaces were selected which repressed the reporter gene to similar levels as the wild-type protein. The leucine-269 alanine-308 core was repeatedly, but not exclusively, selected from the library whilst a diversity of predominantly non-polar residues were selected at positions 267 and 310. These results indicate that L269 and A308 may form a hot spot of interaction and allow an insight into the processes of dimer selectivity and evolution within this family of transcription factors.
Buscar no Google
Base de dados:
MEDLINE
Assunto principal:
Proteínas Repressoras
/
Mutagênese
/
NF-kappa B
/
Escherichia coli
Tipo de estudo:
Clinical_trials
/
Diagnostic_studies
/
Prognostic_studies
/
Screening_studies
Idioma:
En
Ano de publicação:
2001
Tipo de documento:
Article