z-logo
Premium
Effects of small neutral osmolytes on the supercoiling free energy and intrinsic twist of p30δ DNA
Author(s) -
Rangel David P.,
Sucato Christopher A.,
Spink Charles H.,
Fujimoto Bryant S.,
Schurr J. Michael
Publication year - 2004
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.20111
Subject(s) - chemistry , ethylene glycol , crystallography , equilibrium constant , twist , thermodynamics , physics , geometry , organic chemistry , mathematics
Both theory and experiments are employed to investigate the effects of small neutral osmolytes on the average intrinsic twist ( l 0 ), the torsion and bending elastic constants, and the twist energy parameter ( E T ) that governs the supercoiling free energy. The experimental data for ethylene glycol and acetamide at 37°C suggest, and are interpreted in terms of, a model wherein the DNA exhibits an equilibrium between two distinct conformational states that possess different numbers of bound water molecules and exhibit different intrinsic twists and torsion and bending elastic constants. Expressions are derived to relate the effective E T and l 0 to the equilibrium constant, water activity ( a w ), and number ( n ) of bound water molecules released per cooperative domain undergoing the two‐state transition. The variations of l 0 and E T with −ln( a w ) are similar for acetamide and ethylene glycol at 37°C. Fitting the theory to those data yields the range n = 103–125 for ethylene glycol and n = 71–113 for acetamide, depending on the assumed value of E T for the dehydrated state. The cooperative domain size of the two‐state transition is estimated to exceed about 25–30 base pairs (bp). Between 0 and 19.4 w/v % ethylene glycol, the torsion elastic constant, measured by time‐resolved fluorescence polarization anisotropy (FPA), increases by 1.37‐fold, whereas the measured E T decreases by 1.15‐fold over that same range. The implied decrease in bending rigidity over that range is by a factor of about 0.7. The variations of l 0 and E T with increasing −ln( a w ) due to added ethylene glycol at 37°C are far smaller than the corresponding variations observed previously at 14 and 15°C. However, at 21°C, upon adding either ethylene glycol or acetamide, l 0 and E T initially decline steeply with increasing −ln( a w ), with slopes possibly comparable to those seen at 14 and 15°C, but then flatten out and follow curves similar to those at 37°C. Possible origins of such mixed behavior are discussed. The effects of betaine at both 37 and 21°C differ qualitatively and quantitatively in various respects from those of ethylene glycol and acetamide. Upon adding sucrose, l 0 initially jumps to higher plateaus at both 37 and 21°C, but its effects on E T cannot be reliably assessed, due to the limited range of −ln( a w ). © 2004 Wiley Periodicals, Inc. Biopolymers, 2004

This content is not available in your region!

Continue researching here.

Having issues? You can contact us here