Ligand-induced conformational changes and conformational dynamics in the solution structure of the lactose repressor protein.
We present here the results of a series of small-angle X-ray scattering studies aimed at understanding the role of conformational changes and structural flexibility in DNA binding and allosteric signaling in a bacterial transcription regulator, lactose repressor protein (LacI). Experiments were designed to detect possible conformational changes that occur when LacI binds either DNA or the inducer IPTG, or both. Our studies included the native LacI dimer of homodimers and a dimeric variant (R3), enabling us to probe conformational changes within the homodimers and distinguish them from those involving changes in the homodimer-homodimer relationships. The scattering data indicate that removal of operator DNA (oDNA) from R3 results in an unfolding and extension of the hinge helix that connects the LacI regulatory and DNA-binding domains. In contrast, only very subtle conformational changes occur in the R3 dimer-oDNA complex upon IPTG binding, indicative of small adjustments in the orientations of domains and/or subdomains within the structure. The binding of IPTG to native (tetrameric) LacI-oDNA complexes also appears to facilitate a modest change in the average homodimer-homodimer disposition. Notably, the crystal structure of the native LacI-oDNA complex differs significantly from the average solution conformation. The solution scattering data are best fit by an ensemble of structures that includes (1) approximately 60% of the V-shaped dimer of homodimers observed in the crystal structure and (2) approximately 40% of molecules with more "open" forms, such as those generated when the homodimers move with respect to each other about the tetramerization domain. In gene regulation, such a flexible LacI would be beneficial for the interaction of its two DNA-binding domains, positioned at the tips of the V, with the required two of three LacI operators needed for full repression.
Taraban
Marc
M
Department of Chemistry, University of Utah, Salt Lake City, UT 84112, USA.
Zhan
Hongli
H
Whitten
Andrew E
AE
Langley
David B
DB
Matthews
Kathleen S
KS
Swint-Kruse
Liskin
L
Trewhella
Jill
J
D000494:Allosteric Regulation; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D004247:DNA / Q000378:metabolism; D019281:Dimerization; D007544:Isopropyl Thiogalactoside / Q000378:metabolism; D007785:Lactose / Q000037:antagonists & inhibitors*; D008024:Ligands; D008954:Models, Biological; D008958:Models, Molecular; D008970:Molecular Weight; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012097:Repressor Proteins / Q000737:chemistry* / Q000378:metabolism*; D053838:Scattering, Small Angle; D013816:Thermodynamics
D001426:Bacterial Proteins; D008024:Ligands; D012097:Repressor Proteins; D007544:Isopropyl Thiogalactoside; D004247:DNA; D007785:Lactose