W. Klein et al., Molecular characterization of the transition state regulator AbrB from Bacillus stearothermophilus, BBA-GENE ST, 1493(1-2), 2000, pp. 82-90
Citations number
32
Categorie Soggetti
Molecular Biology & Genetics
Journal title
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION
The Bacillus subtilis transition state regulator AbrB(su) is a DNA-binding
protein that acts on several genes either as activator, repressor, or preve
nter. However, among genes under its control, neither common binding sites
could be identified nor could the structural features of this broad and spe
cific interaction be elucidated. Attempts to elucidate these interesting fe
atures by crystallizing AbrB(su) have failed so far. Therefore, to solve th
is problem, we focused in this work on identifying an AbrB(su) homologue fr
om Bacillus stearothermophilus. Using a novel method. the entire abrB(st) g
ene of B. stearothermophilus was cloned and sequenced. The gene encodes a 9
5 amino acid protein that shows 77% identity and 85% similarity to the meso
philic B. subtilis protein. A calmodulin binding peptide-tagged fusion of t
he thermophilic gene was constructed for overexpression and efficient affin
ity column purification of the AbrB(st) protein. The purified protein showe
d, after removal of the tag, an oligomerization behavior through hexamer fo
rmation that is essential for its DNA binding activity. (C) 2000 Elsevier S
cience B.V. All rights reserved.