Association of short DNA fragments: Steady state fluorescence polarization study

We have studied aggregation/association of monodisperse DNA fragments (ranging from 30–90 base pairs) by steady‐state fluorescence polarization of intercalculated ethidium. The method of excitation at different wavelengths in the ethidium absorption spectrum provides information about anisotropic twisting and tumbling mobility of the fragments. We find that end‐over‐end tumbling rather than axial spinning and internal twisting motions are affected by aggregation/association. The critical concentration for observing the effects of intermolecular interactions is approximately 5 mg DNA/mL at room temperature, independent of fragment length. Association is favored by low temperature and high (> 10 mM) concentration of Mg2+. From temperature‐and salt‐dependence experiments we infer that the “aggregates” are similar to those observed in a recently discovered DNA sol–gel transition [M. G. Fried and V. A. Bloomfield (1984) Biopolymers 23, 2141–2155]. We also discuss possible arrangements of the fragments within the aggregates and their possible relation to formation of DNA liquid crystals.

[1]  T. Härd,et al.  Anisotropic overall and internal motions of short DNA fragments. , 1986, Nucleic acids research.

[2]  D. Kearns,et al.  Investigation of DNA dynamics and drug-DNA interaction by steady state fluorescence anisotropy. , 1985, Nucleic acids research.

[3]  M. Fried,et al.  DNA gelation in concentrated solutions , 1984, Biopolymers.

[4]  A. Szabó Theory of fluorescence depolarization in macromolecules and membranes , 1984 .

[5]  G. C. Levy,et al.  Effects of intercalated ethidium on DNA conformational dynamics. A reevaluation by 31P and 13C NMR studies , 1984 .

[6]  N. Assa‐Munt,et al.  1H NMR relaxation studies of the hydrogen-bonded imino protons of poly(dA-dT). , 1984, Biochemistry.

[7]  M. Behe,et al.  B to Z transition of double-stranded poly[deoxyguanylyl(3'-5')-5-methyldeoxycytidine] in solution by phosphorus-31 and carbon-13 nuclear magnetic resonance spectroscopy. , 1983, Biochemistry.

[8]  A. Richard Centrifugal field relaxation and ionic strength effects on calf thymus DNA gels , 1983, Biopolymers.

[9]  G. C. Levy,et al.  Spontaneous ordering of DNA. Effects of intermolecular interactions on DNA motional dynamics monitored by 13C and 31P nuclear magnetic resonance spectroscopy. , 1983, The Journal of biological chemistry.

[10]  R. Wartell,et al.  Changes in Raman vibrational bands of calf thymus DNA during the B‐to‐A transition , 1982, Biopolymers.

[11]  R. J. Robbins,et al.  Torsion and bending of nucleic acids studied by subnanosecond time-resolved fluorescence depolarization of intercalated dyes , 1982 .

[12]  M. Barkley,et al.  Fluorescence decay studies of anisotropic rotations of small molecules , 1981 .

[13]  D. Crothers,et al.  Conversion of B DNA between solution and fiber conformations. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[14]  H. Frisch,et al.  Thermodynamics and equilibrium sedimentation analysis of the close approach of DNA molecules and a molecular ordering transition , 1981, Biopolymers.

[15]  D. Barrow,et al.  Light-scattering effects in the measurement of membrane microviscosity with diphenylhexatriene. , 1979, Biophysical journal.

[16]  C. Houssier,et al.  Interaction of ethidium bromide with DNA. Optical and electrooptical study , 1974, Biopolymers.

[17]  Y. Suezaki,et al.  Ultracentrifugal field relaxation behavior of fd‐DNA , 1973 .

[18]  M. Waring,et al.  Complex formation between ethidium bromide and nucleic acids. , 1965, Journal of molecular biology.

[19]  F. Perrin Polarisation de la lumière de fluorescence. Vie moyenne des molécules dans l'etat excité , 1926 .

[20]  Dietrich Stauffer,et al.  Gelation and critical phenomena , 1982 .

[21]  Sam F. Edwards,et al.  Dynamics of rod-like macromolecules in concentrated solution. Part 1 , 1978 .

[22]  H. Mahler,et al.  SOME OBSERVATIONS ON THE HYPOCHROMISM OF DNA. , 1964, Journal of molecular biology.