DNA base pair resolution by single molecule force spectroscopy.
The forces that hold complementary strands of DNA together in a double helix, and the role of base mismatches in these, are examined by single molecule force spectroscopy using an atomic force microscope (AFM). These forces are important when considering the binding of proteins to DNA, since these proteins often mechanically stretch the DNA during their action. In AFM measurement of forces, there is an inherent instrumental limitation that makes it difficult to compare results from different experimental runs. This is circumvented by using an oligonucleotide microarray, which allowed a direct comparison of the forces between perfectly matched short oligonucleotides and those containing a single or double mismatch. Through this greatly increased sensitivity, the force contribution of a single AT base pair was derived. The results indicate that the contribution to forces from the stacking interactions is more important than that from hydrogen bonding.
Sattin
Bernie D
BD
Department of Chemistry, University of Toronto, Toronto, Ontario M5S 3H6, Canada.
Pelling
Andrew E
AE
Goh
M Cynthia
MC
D020137:Base Pair Mismatch; D020029:Base Pairing; D004247:DNA / Q000737:chemistry*; D006860:Hydrogen Bonding; D018625:Microscopy, Atomic Force / Q000379:methods*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D020411:Oligonucleotide Array Sequence Analysis; D013057:Spectrum Analysis / Q000379:methods*
D009838:Oligodeoxyribonucleotides; D004247:DNA