Alexandria Digital Research Library

Electrostatics of single-stranded nucleic acids : force spectroscopy and ion counting experiments

Author:
Jacobson, David R.
Degree Grantor:
University of California, Santa Barbara. Physics
Degree Supervisor:
Philip A. Pincus and Omar A. Saleh
Place of Publication:
[Santa Barbara, Calif.]
Publisher:
University of California, Santa Barbara
Creation Date:
2016
Issued Date:
2016
Topics:
Biophysics and Physics
Keywords:
Magnetic tweezers
Force spectroscopy
Donnan equilibrium
Nucleic acids
Ion atmosphere
Electrostatics
Genres:
Dissertations, Academic and Online resources
Dissertation:
Ph.D.--University of California, Santa Barbara, 2016
Description:

The strong negative charge of nucleic acids adds additional complexity to their behavior, beyond what would be expected for neutral polymers, through the addition of long-range self-repulsive interactions and through the screening of those interactions by ions in solution. Among the techniques that can probe the electrostatic behavior of a charged polymer are single-molecule force spectroscopy, which reports on the elasticity of the polymer (i.e., extension as a function of applied force), and ion counting, which reports on the stoichiometry of positive and negative ions constituting the polymer's "ion atmosphere". While extensive studies of both types have been made of double-stranded DNA, a strongly charged and moderately stiff polymer, less is known about the single-stranded nucleic acids (ssNAs), which are also strongly charged but are comparatively flexible.

In my dissertation, I use single-molecule magnetic tweezers to measure the elasticity of ssRNA as a function of monovalent and divalent salt concentration under 0.1--10 pN applied force and observe behavior characteristic of flexible polyelectrolytes. I then extend the measurements up to 100 pN to probe the intermediate-force domain connecting behavior dominated by electrostatics and by the chemical structure of the backbone. These new data allow the critical testing of a wormlike chain-derived model treating electrostatics through a mean-field, salt-dependent tension. This model is shown to quantitatively account for the data and to bridge the established behaviors at lower and higher force. I also quantify the ion atmosphere stoichiometry of both ssDNA and ssRNA using three complementary experimental techniques, including both bulk and single-molecule approaches. These results are compared with Poisson-Boltzmann models incorporating varying degrees of structural complexity, demonstrating the inherently short-ranged (i.e., sub-Debye screening length) nature of nucleic acid-ion interactions.

Physical Description:
1 online resource (215 pages)
Format:
Text
Collection(s):
UCSB electronic theses and dissertations
ARK:
ark:/48907/f33t9hcj
ISBN:
9781369576139
Catalog System Number:
990047511930203776
Rights:
Inc.icon only.dark In Copyright
Copyright Holder:
David Jacobson
File Description
Access: Public access
Jacobson_ucsb_0035D_13265.pdf pdf (Portable Document Format)