title: Structure of the Tet repressor and Tet repressor-operator complexes in solution from electrooptical measurements and hydrodynamic simulations.
abstract: The Tet repressor protein and tet operator DNA fragments and their complexes have been analyzed by electrooptical procedures. The protein shows a positive linear dichroism at 280 nm, a negative linear dichroism at 248 nm, and a strong permanent dipole moment of 3.5 X 10(-27) C m, which is independent of the salt concentration within experimental accuracy. Its rotation time constant of 40 ns indicates an elongated structure, which is consistent with a prolate ellipsoid of 100 A for the long axis and 40 A for the short axis. The time constant can also be fitted by a cylinder of length 78 A and diameter 37 A, which is consistent with nuclease protection data reported on repressor-operator complexes, if the cylinder axis is aligned parallel to the DNA axis. Addition of tetracycline induces changes of the limit dichroism but very little change of the rotation time constant. The rotation time constants observed for the operator DNA fragments show some deviations from the values expected from their contour length; however, these deviations remain relatively small. Formation of repressor-operator complexes leads to some increase of the DNA rotation time constants. Simulations by bead models demonstrate that these time constants can be explained without any major change of the hydrodynamic dimension of the components. The data for the complexes are fitted by bead models with smooth bending of the DNA corresponding to a radius of curvature of 500 A, but at the given accuracy, we cannot rule out that the DNA in the complex remains straight or is bent to a smaller radius of approximately 400 A. Thus, binding of the Tet repressor, which is a helix-turn-helix protein as judged from its sequence, to its operator seems to induce minor bending but does not induce strong bending of the DNA double helix.
authors.affiliation: Max-Planck-Institut für biophysikalische Chemie, Göttingen, FRG.
authors.forename: D
authors.initials: D
authors.lastname: Porschke
authors.forename: K
authors.initials: K
authors.lastname: Tovar
authors.forename: J
authors.initials: J
authors.lastname: Antosiewicz
mesh_terms: D004251:DNA Transposable Elements; D004563:Electrochemistry; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D011815:R Factors; D012097:Repressor Proteins / Q000378:metabolism*; D012996:Solutions; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D012097:Repressor Proteins; D012996:Solutions; D014157:Transcription Factors; C034811:tetracycline resistance-encoding transposon repressor protein