Superhelical DNA as a preferential binding target of 14-3-3γ protein
Language English Country England, Great Britain Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Escherichia coli genetics MeSH
- HCT116 Cells MeSH
- Cloning, Molecular MeSH
- Binding, Competitive MeSH
- DNA, Cruciform genetics metabolism MeSH
- Humans MeSH
- Plasmids genetics MeSH
- 14-3-3 Proteins genetics metabolism MeSH
- Recombinant Fusion Proteins genetics metabolism MeSH
- DNA Replication genetics MeSH
- Electrophoretic Mobility Shift Assay MeSH
- DNA, Superhelical genetics metabolism MeSH
- Protein Binding MeSH
- Binding Sites MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- DNA, Cruciform MeSH
- 14-3-3 Proteins MeSH
- Recombinant Fusion Proteins MeSH
- DNA, Superhelical MeSH
The 14-3-3 protein family is a highly conserved and widely distributed group of proteins consisting of multiple isoforms in eukaryotes. Ubiquitously expressed, 14-3-3 proteins play key roles in DNA replication, cell cycle regulation, and apoptosis. The function of 14-3-3 proteins is mediated by interaction with a large number of other proteins and with DNA. It has been demonstrated that 14-3-3γ protein binds strongly to cruciform structures and is crucial for initiating replication. In this study, we analyzed DNA binding properties of the 14-3-3γ isoform to linear and supercoiled DNA. We demonstrate that 14-3-3γ protein binds strongly to long DNA targets, as evidenced by electrophoretic mobility shift assay on agarose gels. Binding of 14-3-3γ to DNA target results in the appearance of blurry, retarded DNA bands. Competition experiments with linear and supercoiled DNA on magnetic beads show very strong preference for supercoiled DNA. We also show by confocal microscopy that 14-3-3 protein in the HCT-116 cell line is co-localized with DNA cruciforms. This implies a role for the 14-3-3γ protein in its binding to local DNA structures which are stabilized by DNA supercoiling.
References provided by Crossref.org
Interaction of Proteins with Inverted Repeats and Cruciform Structures in Nucleic Acids
