Distorted DNA structures induced by HMGB2 possess a high affinity for HMGB2.
HMGB2 (HMG2) protein binds with DNA duplex in a sequence-nonspecific manner, then bends and unwinds the DNA. In DNA cyclization analyses for the bending activity of HMGB2, two unidentified bands, denoted alpha and beta, were observed in addition to monomer circular DNA (1C) on the gel. Re-electrophoresis and proteinase K digestion revealed that alpha and beta are complexes of circularized probe DNA (seeming 1C) with HMGB2 (K(d) approximately 10(-10) M). The DNA components of alpha and beta (alpha- and beta-DNA) showed higher affinities to HMGB2 than did the linear probe DNA (K(d) approximately 10(-7) M). The DNAs have distorted structures containing partial single-stranded regions. Nicked circular molecules presumably due to severe DNA distortion by HMGB2 were observed in alpha- and beta-DNA, in addition to closed circular double-stranded molecules. The alpha and beta bands were not formed in the presence of sole DNA binding regions which are necessary for DNA bending, indicating that the acidic C-tail in the HMGB2 molecule is necessary for inducing the peculiar distorted structures of higher affinity to HMGB2. HMGB2 binds with linker DNA and/or the entry and exit of nucleosomes fixed at both ends likewise mini-circles similar to alpha-DNA and beta-DNA. Thus, the distorted structures present in alpha-DNA and beta-DNA should be important in considering the functional mechanisms in which HMGB2 participates.
Nakamura
Yasuyuki
Y
Department of Biological Science and Technology, Science University of Tokyo Yamazaki, Noda, Chiba 278-8510, Japan.
Shimizu
Mitsuhiro
M
Yoshida
Michiteru
M
D000818:Animals; D003500:Cyclization; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D004270:DNA, Circular / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D024742:HMG-Box Domains / Q000502:physiology; D024261:HMGB2 Protein / Q000378:metabolism*; D015335:Molecular Probes; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D013552:Swine
D004270:DNA, Circular; D004268:DNA-Binding Proteins; D024261:HMGB2 Protein; D015335:Molecular Probes; D004247:DNA; D003851:Deoxyribonucleases; D004264:DNA Topoisomerases, Type I