We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Synchrotron Radiation Circular Dichroism, a New Tool to Probe Interactions between Nucleic Acids Involved in the Control of ColE1-Type Plasmid Replication.
- Authors
Wien, Frank; Kubiak, Krzysztof; Turbant, Florian; Mosca, Kevin; Węgrzyn, Grzegorz; Arluison, Véronique
- Abstract
Featured Application: Our results indicate previously unknown macromolecular interplays in the regulation of ColE1-like plasmid DNA replication, which should be important in further studies on combating bacterial antibiotic resistance through controlling the propagation and spreading of plasmids as carriers of antibiotic resistance genes. New insights on this system are brought by synchrotron radiation-based circular dichroism (SRCD), which gives access to an extended spectral range down to the vacuum UV (vUV) range. Only the synchrotron light source provides intensities of the beam below 190 nm, which allows a precise determination of the nucleic acid conformation. Hfq is a bacterial master regulator which promotes the pairing of nucleic acids. Due to the high molecular weight of the complexes formed between nucleic acids and the amyloid form of the protein, it is difficult to analyze solely by a gel shift assay the complexes formed, as they all migrate at the same position in the gel. In addition, precise kinetics measurements are not possible using a gel shift assay. Here, we used a synchrotron-based biophysical approach, synchrotron radiation circular dichroism (SRCD), to probe the interaction of the Escherichia coli Hfq C-terminal amyloid region with nucleic acids involved in the control of ColE1-like plasmid replication. We observed that this C-terminal region of Hfq has an unexpected and significant effect on the annealing of nucleic acids involved in this process and, more importantly, on their alignment. Functional consequences of this newly discovered property of the Hfq amyloid region are discussed in terms of the biological significance of Hfq in the ColE1-type plasmid replication process and antibiotic resistance.
- Subjects
SYNCHROTRON radiation; CIRCULAR dichroism; PLASMIDS; DNA replication; NUCLEIC acids; MOLECULAR weights; LIGHT sources; AMYLOID beta-protein
- Publication
Applied Sciences (2076-3417), 2022, Vol 12, Issue 5, p2639
- ISSN
2076-3417
- Publication type
Article
- DOI
10.3390/app12052639