Quantitative determination of size and shape of surface-bound DNA using an acoustic wave sensor.
DNA bending plays a significant role in many biological processes, such as gene regulation, DNA replication, and chromosomal packing. Understanding how such processes take place and how they can, in turn, be regulated by artificial agents for individual oriented therapies is of importance to both biology and medicine. In this work, we describe the application of an acoustic wave device for characterizing the conformation of DNA molecules tethered to the device surface via a biotin-neutravidin interaction. The acoustic energy dissipation per unit mass observed upon DNA binding is directly related to DNA intrinsic viscosity, providing quantitative information on the size and shape of the tethered molecules. The validity of the above approach was verified by showing that the predesigned geometries of model double-stranded and triple-helix DNA molecules could be quantitatively distinguished: the resolution of the acoustic measurements is sufficient to allow discrimination between same size DNA carrying a bent at different positions along the chain. Furthermore, the significance of this analysis to the study of biologically relevant systems is shown during the evaluation of DNA conformational change upon protein (histone) binding.
Tsortos
Achilleas
A
Institute of Molecular Biology and Biotechnology, Foundation for Research & Technology Hellas, Vassilika Vouton, 71110 Heraklion, Greece.
Papadakis
George
G
Mitsakakis
Konstantinos
K
Melzak
Kathryn A
KA
Gizeli
Electra
E
D000162:Acoustics / Q000295:instrumentation*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D004247:DNA / Q000737:chemistry* / Q000648:ultrastructure*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008839:Microelectrodes; D008970:Molecular Weight; D014159:Transducers
D004247:DNA