DNA bending induced by high mobility group proteins studied by fluorescence resonance energy transfer.
The HMG domains of the chromosomal high mobility group proteins homologous to the vertebrate HMG1 and HMG2 proteins preferentially recognize distorted DNA structures. DNA binding also induces a substantial bend. Using fluorescence resonance energy transfer (FRET), we have determined the changes in the end-to-end distance consequent on the binding of selected insect counterparts of HMG1 to two DNA fragments, one of 18 bp containing a single dA(2) bulge and a second of 27 bp with two dA(2) bulges. The observed changes are consistent with overall bend angles for the complex of the single HMG domain with one bulge and of two domains with two bulges of approximately 90-100 degrees and approximately 180-200 degrees, respectively. The former value contrasts with an inferred value of 150 degrees reported by Heyduk et al. (1) for the bend induced by a single domain. We also observe that the induced bend angle is unaffected by the presence of the C-terminal acidic region. The DNA bend of approximately 95 degrees observed in the HMG domain complexes is similar in magnitude to that induced by the TATA-binding protein (80 degrees), each monomeric unit of the integration host factor (80 degrees), and the LEF-1 HMG domain (107 degrees). We suggest this value may represent a steric limitation on the extent of DNA bending induced by a single DNA-binding motif.
Lorenz
M
M
Institute for Molecular Biotechnology, Jena, Germany.
Hillisch
A
A
Payet
D
D
Buttinelli
M
M
Travers
A
A
Diekmann
S
S
D000595:Amino Acid Sequence; D000818:Animals; D002683:Chironomidae; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D004270:DNA, Circular / Q000737:chemistry / Q000378:metabolism; D004330:Drosophila; D004735:Energy Transfer; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006609:High Mobility Group Proteins / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D012235:Rhodamines / Q000737:chemistry; D012441:Saccharomyces cerevisiae; D013050:Spectrometry, Fluorescence / Q000379:methods
C106141:6-carboxytetramethylrhodamine; D004270:DNA, Circular; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006609:High Mobility Group Proteins; D012235:Rhodamines; C024098:6-carboxyfluorescein; D004247:DNA