Intrinsic bending in GGCC tracts as probed by fluorescence resonance energy transfer.
Double-stranded oligonucleotides containing the sequence 5'-GGCC-3' can be intrinsically bent, according to X-ray crystallography and gel electrophoresis mobility studies. We have performed fluorescence resonance energy transfer (FRET) experiments with dye-labeled oligonucleotides to further investigate the solution structure of this sequence. We find that 5'-GGCC-3'-containing oligonucleotides bring 5'-attached donor and acceptor dyes much closer together than a comparable "straight" sequence that contains 5'-GCGC-3'. The bend angle for the 5'-GGCC-3' sequence is estimated to be approximately 70 degrees, much larger than the crystallographically observed 23 degrees kink but in agreement with other FRET work. In contrast to gel electrophoresis studies, divalent metal ions do not promote increased kinking in 5'-GGCC-3' above background as judged by FRET. Thus, sequence-dependent structural effects in DNA may be a complicating feature of FRET experiments.
Wildeson
J
J
Department of Chemistry and Biochemistry, University of South Carolina, Columbia, South Carolina 29208, USA.
Murphy
C J
CJ
D002851:Chromatography, High Pressure Liquid; D018360:Crystallography, X-Ray; D008274:Magnesium / Q000737:chemistry; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods*; D015032:Zinc / Q000737:chemistry
D009841:Oligonucleotides; D008274:Magnesium; D015032:Zinc