Structural perturbations induced in linear and circular DNA by the architectural protein HU from Bacillus stearothermophilus.
HU is a small DNA-binding protein of eubacteria that is believed to induce or stabilize bending of the double helix and mediate nucleoid compaction in vivo. Although HU does not bind preferentially to specific DNA sequences, it is known to have high affinity for DNA sites containing structural anomalies, such as unpaired or mismatched bases, nicks, and four-way junctions. We have employed Raman spectroscopy to further investigate the structural basis of HU-DNA recognition in solution. Experiments were carried out on the homodimeric HU protein of Bacillus stearothermophilus (HUBst) and a 222-bp DNA fragment, which was isolated in linear (DNA(L222)) and circular (DNA(C222)) forms. In the absence of bound HUBst the Raman signatures of DNA(L222) and DNA(C222) are nearly superimposable, indicating that circularization produces no substantial change in the local B-DNA conformation. Conversely, the Raman signatures of DNA(L222) and DNA(C222) are perturbed significantly and specifically by HUBst binding. The HUBst-induced perturbations are markedly greater for the circularized DNA target. These results support an opportunistic molecular mechanism, in which HU binding is facilitated by intrinsic nonlinearity or flexibility in the DNA target. We propose that DNA segments which are bent or predisposed toward bending provide the high-affinity sites for HU attachment and nucleoid condensation. This model is consistent with the wide range of DNA bending angles reported in crystal structures of HU-DNA complexes.
Benevides
James M
JM
School of Biological Sciences, University of Missouri-Kansas City, 5100 Rockhill Road, Kansas City, Missouri 64110-2499, USA.
Serban
Doinita
D
Thomas
George J
GJ
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D004270:DNA, Circular / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D001411:Geobacillus stearothermophilus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018625:Microscopy, Atomic Force; D009690:Nucleic Acid Conformation; D013059:Spectrum Analysis, Raman
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004270:DNA, Circular; D004268:DNA-Binding Proteins; C040330:histone-like protein HU, bacteria