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Conformational Changes in Single-Strand DNA as a Function of Temperature by SANS

J. Zhou ^*, S. K. Gregurick ^*, S. Krueger  and F. P. Schwarz 

^* Department of Chemistry and Biochemistry, University of Maryland, Baltimore, Maryland; NIST Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland; and Center for Advanced Research in Biotechnology, National Institute of Standards and Technology, Rockville, Maryland

Correspondence: Address reprint requests to F. P. Schwarz, Tel.: 301-738-6219; E-mail: frederick.schwarz{at}nist.gov.

Small-angle neutron scattering (SANS) measurements were performed on a solution of single-strand DNA, 5'-ATGCTGATGC-3', in sodium phosphate buffer solution at 10°C temperature increments from 25°C to 80°C. Cylindrical, helical, and random coil shape models were fitted to the SANS measurements at each temperature. All the shapes exhibited an expansion in the diameter direction causing a slightly shortened pitch from 25°C to 43°C, an expansion in the pitch direction with a slight decrease in the diameter from 43°C to 53°C, and finally a dramatic increase in the pitch and diameter from 53°C to 80°C. Differential scanning calorimeter scans of the sequence in solution exhibited a reversible two-state transition profile with a transition temperature of 47.5 ± 0.5°C, the midpoint of the conformational changes observed in the SANS measurements, and a calorimetric transition enthalpy of 60 ± 3 kJ mol^–1 that indicates a broad transition as is observed in the SANS measurements. A transition temperature of 47 ± 1°C was also obtained from ultraviolet optical density measurements of strand melting scans of the single-strand DNA. This transition corresponds to unstacking of the bases of the sequence and is responsible for the thermodynamic discrepancy between its binding stability to its complementary sequence determined directly at ambient temperatures and determined from extrapolated values of the melting of the duplex at high temperature.

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