The binding motif recognized by HU on both nicked and cruciform DNA

• Published in: The EMBO journal 1999-10, Vol.18 (19), p.5434-5444
• Main Authors: Kamashev, D., Balandina, A., Rouviere-Yaniv, J.
• Format: Article
• Language: English
• Subjects: Base Sequence; Binding Sites; Brassicaceae - genetics; Deoxyribonucleic acid; DNA; DNA containing a nick; DNA Footprinting; DNA junction; DNA structural binding motif; DNA, Plant - chemistry; DNA, Plant - metabolism; heterodimeric protein; histone-like protein; Hydroxyurea - metabolism; Nucleic Acid Conformation; Phenanthrolines - chemistry
• ISSN: 0261-4189; 1460-2075; 1460-2075
• DOI: 10.1093/emboj/18.19.5434

The heterodimeric HU protein, highly conserved in bacteria and involved in transposition, recombination, DNA repair, etc., shares similarity with histones and HMGs. HU, which binds DNA with low affinity and without sequence specificity, binds strongly and specifically to DNA junctions and DNA containing single‐strand breaks. The fine structure of these specific complexes was studied by footprinting and HU chemically converted into nucleases. The positioning of HUαβ on nicked DNA is asymmetrical and specifically oriented: the β‐arm binds the area surrounding the break whereas the α‐arm lies on the 3′ DNA branch. This positioning necessitates a pronounced bend in the DNA at the discontinuous point, which was estimated by circular permutation assay to be 65°. At junctions, HU is similarly asymmetrically positioned in an identical orientation: the junction point plays the role of the discontinuous point in the nicked DNA. The HU binding motif present in both structures is a pair of inclined DNA helices.