Analysis of the NF-κB p50 dimer interface by diversity screening

Hart, Darren J, Speight, Robert E., Sutherland, John D, & Blackburn, Jonathan M (2001) Analysis of the NF-κB p50 dimer interface by diversity screening. Journal of Molecular Biology, 310(3), pp. 563-575.

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Abstract

An in vivo screen has been devised for NF-κB 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.

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ID Code: 70989
Item Type: Journal Article
Refereed: Yes
Additional URLs:
Keywords: NF-κB, homodimer, random mutagenesis, dimer interface, DNA binding
DOI: 10.1006/jmbi.2001.4724
ISSN: 0022-2836
Divisions: Current > Schools > School of Chemistry, Physics & Mechanical Engineering
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Deposited On: 04 May 2014 23:21
Last Modified: 25 Jun 2014 22:47

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