*********************************
There is now a CONTENT FREEZE for Mercury while we switch to a new platform. It began on Friday, March 10 at 6pm and will end on Wednesday, March 15 at noon. No new content can be created during this time, but all material in the system as of the beginning of the freeze will be migrated to the new platform, including users and groups. Functionally the new site is identical to the old one. webteam@gatech.edu
*********************************
Title: Hybridization and dehybridization of short oligonucleotides subject to weak tension
Name: Derek Hart
Abstract: DNA strand separation and formation underlie our most fundamental genetic processes. One of the most powerful tools for exploring these binding and unbinding reactions is single-molecule force spectroscopy. This method applies tension to a single DNA strand and observes the change in its extension or kinetics with a microscope. However, these experiments typically study longer DNA molecules (< 10 base pairs) subject to higher forces (>10 piconewtons); hence, the behavior of short DNA subject to weak forces is not well understood. In particular, it is not clear whether statistical chain models ordinarily used to explain force spectroscopy data are applicable at these small scales. To remedy this, we focused solely on the kinetics and structure of short DNA duplexes (< 10 bp) subject to very weak forces (< 7 piconewtons). For this purpose, we used two tools: an experimental technique called single-molecule fluorescence resonance energy transfer (smFRET), and the coarse-grained DNA simulation code oxDNA. The experiments implemented a simple, high-throughput DNA assay, dubbed “DNA bows”, which exploit the bending rigidity of DNA to exert very weak forces on short DNA strands. With this method, we demonstrate that weak force accelerates the hybridization and dehybridization of short oligonucleotides from 2 to 6 pN, contradicting the predictions of simple chain models. We next used oxDNA to investigate how the extension of short DNA changes with force throughout its binding and unbinding transitions. Regardless of force, we find that DNA extension is higher in its transition state than in its bound state, in agreement with our experiments. We also find that extension is a poor reaction coordinate at 3 piconewtons and below. These results establish the nature of DNA duplex transitions at the force and length scales relevant to DNA biology as well as emerging DNA nanotechnologies.
Location: Howey N110
Committee Members:
Harold Kim, PhD (advisor)
Kurt Weisenfeld, PhD
Kirill Lobachev, PhD
Loren Williams, PhD
James Gumbart, PhD
