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An intrinsically disordered transcription activation domain increases the DNA binding affinity and reduces the specificity of NFκB p50/RelA
- Source :
- The Journal of biological chemistry, vol 298, iss 9
- Publication Year :
- 2022
-
Abstract
- Many transcription factors contain intrinsically disordered transcription activation domains (TADs), which mediate interactions with coactivators to activate transcription. Historically, DNA-binding domains and TADs have been considered as modular units, but recent studies have shown that TADs can influence DNA binding. Whether these results can be generalized to more TADs is not clear. Here, we biophysically characterized the NFκB p50/RelA heterodimer including the RelA TAD and investigated the TAD's influence on NFκB-DNA interactions. In solution, we show the RelA TAD is disordered but compact, with helical tendency in two regions that interact with coactivators. We determined that the presence of the TAD increased the stoichiometry of NFκB-DNA complexes containing promoter DNA sequences with tandem κB recognition motifs by promoting the binding of NFκB dimers in excess of the number of κB sites. In addition, we measured the binding affinity of p50/RelA for DNA containing tandem κB sites and single κB sites. While the presence of the TAD enhanced the binding affinity of p50/RelA for all κB sequences tested, it also increased the affinity for nonspecific DNA sequences by over 10-fold, leading to an overall decrease in specificity for κB DNA sequences. In contrast, previous studies have generally reported that TADs decrease DNA-binding affinity and increase sequence specificity. Our results reveal a novel function of the RelA TAD in promoting binding to nonconsensus DNA, which sheds light on previous observations of extensive nonconsensus DNA binding by NFκB invivo in response to strong inflammatory signals.
- Subjects :
- Transcriptional Activation
Biochemistry & Molecular Biology
cooperativity
1.1 Normal biological development and functioning
Biochemistry
Medical and Health Sciences
Protein Domains
Underpinning research
Genetics
Molecular Biology
structural model
Base Sequence
DNA-protein interaction
Transcription Factor RelA
NF-kappa B p50 Subunit
Cell Biology
DNA
Biological Sciences
intrinsically disordered protein
hydrogen-deuterium exchange
small-angle X-ray scattering
Chemical Sciences
Generic health relevance
Protein Multimerization
transcription
NF-kB transcription factor
Protein Binding
Subjects
Details
- ISSN :
- 1083351X
- Volume :
- 298
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- The Journal of biological chemistry
- Accession number :
- edsair.doi.dedup.....20304361ba5d0df802ef2b7d18b255af