Changes in DNA superhelical density monitored by polarized light scattering.
Linear and circular lambda-DNA at different ethidium bromide concentrations have been studied by means of polarized light scattering, namely the S14, S34, S33 and S13 elements of Mueller matrix. While S33 at low angle appears well correlated with the total light scattering evaluated by optical density measurements at 632.8 nm for linear and circular DNA of the same mass, the magnitude and slope of the S14, S34 and S13 signals display significant changes for the circular lambda-DNA depending on the degree of negative superhelical density as induced by the different ethidium bromide concentrations. At the same time, for linear lambda-DNA the signal remains invariant, making explicit for the differential scattering of polarized light the possibility to obtain additional information by its angular dependence. Strikingly also the effect of 0.2% glutaraldehyde versus ethanol fixation on the native lambda-DNA structural properties appears to confirm earlier findings by other well-established probes. Results are discussed in terms of first physical principles and of their potential bearings towards our understanding of the mechanism controlling gene expression.
Nicolini
C
C
Institute of Biophysics, University of Genova, Italy.
Diaspro
A
A
Bertolotto
M
M
Facci
P
P
Vergani
L
L
D010582:Bacteriophage lambda; D004270:DNA, Circular / Q000737:chemistry*; D004278:DNA, Superhelical / Q000737:chemistry* / Q000187:drug effects; D004279:DNA, Viral / Q000737:chemistry* / Q000187:drug effects; D005976:Glutaral / Q000494:pharmacology; D007700:Kinetics; D008027:Light; D009690:Nucleic Acid Conformation; D012542:Scattering, Radiation
D004270:DNA, Circular; D004278:DNA, Superhelical; D004279:DNA, Viral; D005976:Glutaral