DNA-binding properties of the tandem HMG boxes of high-mobility-group protein 1 (HMG1).
High-mobility-group protein 1 (HMG1) is a conserved chromosomal protein with two homologous DNA-binding HMG-box domains, A and B, linked by a short basic region to an acidic carboxy-terminal tail. NMR spectroscopy on the free didomain (AB) shows that the two HMG boxes do not interact. The didomain has a higher affinity for all DNA substrates tested than single HMG-box domains and has a significantly higher ability to distort DNA by bending and supercoiling. The interaction of the didomain with DNA is stabilized by the presence of the basic region (approximately 20 residues, 9 of which are Lys) that links the second HMG box to the acidic tail in intact HMG1; this may be, at least in part, why this region also enhances supercoiling of relaxed circular DNA by the didomain and circularization of short DNA fragments (in the presence of ligase). Competition assays suggest significantly different structure-specific preferences of single and tandem HMG boxes for four-way junction and supercoiled plasmid DNA. Binding to supercoiled DNA appears to be promoted by protein oligomerization, which is pronounced for the didomains. Electron microscopy suggests that the oligomers are globular aggregates, associated with DNA looping. One box versus two (or several) is likely to be an important determinant of the properties of (non-sequence specific) HMG-box proteins.
Grasser
K D
KD
Cambridge Centre for Molecular Recognition and Department of Biochemistry, University of Cambridge, UK.
Teo
S H
SH
Lee
K B
KB
Broadhurst
R W
RW
Rees
C
C
Hardman
C H
CH
Thomas
J O
JO
D001665:Binding Sites; D001667:Binding, Competitive; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D017931:DNA Primers; D004278:DNA, Superhelical / Q000737:chemistry / Q000378:metabolism; D006609:High Mobility Group Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008854:Microscopy, Electron; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000737:chemistry; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism
D017931:DNA Primers; D004278:DNA, Superhelical; D006609:High Mobility Group Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D004247:DNA