Helmholtz Gemeinschaft


Recognition of T-rich single-stranded DNA by the cold shock protein Bs-CspB in solution

[thumbnail of 13632oa.pdf] PDF - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader

Item Type:Article
Title:Recognition of T-rich single-stranded DNA by the cold shock protein Bs-CspB in solution
Creators Name:Zeeb, M., Max, K.E.A., Weininger, U., Loew, C., Sticht, H. and Balbach, J.
Abstract:Cold shock proteins (CSP) belong to the family of single-stranded nucleic acid binding proteins with OB-fold. CSP are believed to function as 'RNA chaperones' and during anti-termination. We determined the solution structure of Bs-CspB bound to the single-stranded DNA (ssDNA) fragment heptathymidine (dT7) by NMR spectroscopy. Bs-CspB reveals an almost invariant conformation when bound to dT7 with only minor reorientations in loop {beta}1-{beta}2 and {beta}3-{beta}4 and of few aromatic side chains involved in base stacking. Binding studies of protein variants and mutated ssDNA demonstrated that Bs-CspB associates with ssDNA at almost diffusion controlled rates and low sequence specificity consistent with its biological function. A variation of the ssDNA affinity is accomplished solely by changes of the dissociation rate. 15N NMR relaxation and H/D exchange experiments revealed that binding of dT7 increases the stability of Bs-CspB and reduces the sub-nanosecond dynamics of the entire protein and especially of loop {beta}3-{beta}4.
Keywords:Bacillus Subtilis, Bacterial Proteins, Biomolecular Nuclear Magnetic Resonance, DNA Mutational Analysis, Deuterium Exchange Measurement, Kinetics, Molecular Models, Protein Conformation, Single-Stranded DNA, Thermodynamics, Thymidine
Source:Nucleic Acids Research
Publisher:Oxford University Press
Page Range:4561-4571
Official Publication:https://doi.org/10.1093/nar/gkl376
PubMed:View item in PubMed

Repository Staff Only: item control page


Downloads per month over past year

Open Access
MDC Library