The quaternary structure of Tet repressors bound to the Tn10-encoded tet gene control region determined by neutron solution scattering.

AUTOR(ES)

Lederer, H

RESUMO

The spatial arrangement of Tet repressor dimer, both free and in complex with an 80 bp DNA fragment spanning the wild-type Tn10-encoded tet transcriptional control sequence containing a tandem repeat of two operators, has been determined by neutron small-angle scattering. The active, free Tet repressor dimer is an elongated and flat molecule with a maximum dimension of 11 +/- 1.5 mm which can be approximated by an ellipsoid with the half-axes 6 nm, 2.5 nm and 1 nm. The overall conformation undergoes no detectable change when the repressor dimer is bound to a DNA fragment containing a single tet operator. The normal distance between the centre of gravity of the protein and the DNA axis is 3.0 +/- 0.1 nm, indicating that the repressor dimer is mainly located on one side of the DNA. When bound to the wild type tet control DNA, the two repressor dimers have a centre-to-centre distance of 11.0 +/- 0.5 nm. Their minimal distance is 5 +/- 2 nm. Protein-protein contacts via loop formation of the DNA by repressor binding is excluded. The repressors are well separated and have no direct contact. A model is proposed where the two repressor dimers are located on opposite sides of the DNA and the DNA is not strongly bent in the complex.

Documentos Relacionados

• A multifunctional gene (tetR) controls Tn10-encoded tetracycline resistance.
• Correlation of thermodynamic and genetic properties in the Tn10 encoded TET gene control region.
• Tn10-encoded tet repressor can regulate an operator-containing plant promoter.
• Cholesteryl myristate conformation in liquid crystalline mesophases determined by neutron scattering.
• Differential regulation of the Tn10-encoded tetracycline resistance genes tetA and tetR by the tandem tet operators O1 and O2.