| Status | 已發表Published |
| Structural and kinetic study reveal the dynamic of NFκB:DNA recognition | |
Pan, W. ; Meshcheryakov, V.; Wang, V. Y.-F. 
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| 2021-07-23 | |
| Source Publication | The 7th Macau Symposium on Biomedical Sciences Abstract Book
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| Abstract | NFκB is a family of transcription factors, consist of five subunits: RelA (p65), RelB, c-Rel, p50 and p52. The binding of NFκB to its specific binding sites on promotor or enhancer of genomic DNA is a critical event in regulating its target gene expression. NFκB bind to consensus DNA 5’-GGGNNNNNCC-3’, known as κB sites or κB DNA. The κB DNAs are pseudo symmetric with two half-sites; NFκB always forms and functions as a dimer. Each NFκB monomer bind to one half-site of κB DNA. The crystal structures of NFκB:DNA complex indicated that the G and C base pairs at the 5’ and 3’ terminus are the most important in NFκB:DNA recognition; the central base pair (bp) of κB DNA is not directly contacted by NFκB. However, previous study of our group found out that the identity of central bp of κB sites plays an important role in transcriptional specificity. To determine the molecular mechanism behind this effect, we crystallized the complex of NFκB p52 homodimer and G/C-centric κB DNA and the X-ray crystal structure showed that the overall structure was similar to other known NFκB:DNA structures, but the G/C-centric DNA shows wider minor groove than regular B-form DNA in the central part of κB sites. We analyzed the thermodynamic and binding kinetic of the p52 homodimer with various κB DNAs. The thermodynamic analyses by isothermal titration calorimetry (ITC) revealed that the interaction of p52 homodimer with G/C-centric DNA was driven by entropy, whereas the A/T-centric DNA was bound with similar affinity due to larger change in enthalpy. The binding kinetic analyzed by biolayer interferometry (BLI) showed that the association rate and dissociation rate of G/C-centric κB DNA binding to p52 homodimer is slightly higher than A/T-centric κB DNA. These suggest the mechanism of p52 binding to different κB DNAs was not identical. The p52 homodimer required of its cofactor Bcl3 to activate the transcription, so we also tried to determine the structure of the p52:Bcl3:DNA ternary complex by cryo-EM, and the preliminary result showed dispersed particles of approximately 150Å diameter for now, we will put our effort on promoting the resolution. |
| Keyword | Crystal structure kinetics thermodynamics |
| Language | 英語English |
| The Source to Article | PB_Publication |
| PUB ID | 58700 |
| Document Type | Conference paper |
| Collection | DEPARTMENT OF BIOMEDICAL SCIENCES |
| Corresponding Author | Wang, V. Y.-F. |
| Recommended Citation GB/T 7714 | Pan, W.,Meshcheryakov, V.,Wang, V. Y.-F.. Structural and kinetic study reveal the dynamic of NFκB:DNA recognition[C], 2021. |
| APA | Pan, W.., Meshcheryakov, V.., & Wang, V. Y.-F. (2021). Structural and kinetic study reveal the dynamic of NFκB:DNA recognition. The 7th Macau Symposium on Biomedical Sciences Abstract Book. |
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