2018
DOI: 10.1021/acs.biochem.8b00158
|View full text |Cite
|
Sign up to set email alerts
|

Protein Cofactors Are Essential for High-Affinity DNA Binding by the Nuclear Factor κB RelA Subunit

Abstract: Transcription activator proteins typically contain two functional domains: a DNA binding domain (DBD) that binds to DNA with sequence specificity and an activation domain (AD) whose established function is to recruit RNA polymerase. In this report, we show that purified recombinant nuclear factor κB (NF-κB) RelA dimers bind specific κB DNA sites with an affinity significantly lower than that of the same dimers from nuclear extracts of activated cells, suggesting that additional nuclear cofactors might facilita… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
1
1
1
1

Citation Types

1
9
0

Year Published

2019
2019
2024
2024

Publication Types

Select...
6
1
1

Relationship

1
7

Authors

Journals

citations
Cited by 17 publications
(10 citation statements)
references
References 50 publications
1
9
0
Order By: Relevance
“…Furthermore, the binding affinity of TFs to DNA can be subject to modulation by recruitment of other factors, e.g., low-affinity can change into high-affinity via cofactor binding [ 28 ]. For example, recombinant RelA dimers bind to nuclear factor kappa B (NFκB) DNA sites; however, when another TF such as p53 is added, RelA DNA binding affinity increases up to 4-fold [ 29 ]. If a TF binds DNA, this does not necessarily result in gene expression; this can be threshold-dependent, i.e., binding of low levels of TF does not induce gene expression, but binding of TFs above a certain threshold concentration activates gene expression, creating a switch-like effect ( Figure 1 B).…”
Section: Transcription Factorsmentioning
confidence: 99%
“…Furthermore, the binding affinity of TFs to DNA can be subject to modulation by recruitment of other factors, e.g., low-affinity can change into high-affinity via cofactor binding [ 28 ]. For example, recombinant RelA dimers bind to nuclear factor kappa B (NFκB) DNA sites; however, when another TF such as p53 is added, RelA DNA binding affinity increases up to 4-fold [ 29 ]. If a TF binds DNA, this does not necessarily result in gene expression; this can be threshold-dependent, i.e., binding of low levels of TF does not induce gene expression, but binding of TFs above a certain threshold concentration activates gene expression, creating a switch-like effect ( Figure 1 B).…”
Section: Transcription Factorsmentioning
confidence: 99%
“…Furthermore, NEIL2 was found to directly interact with the DNA-binding domain of RelA, a component J o u r n a l P r e -p r o o f of the NF-κB dimer, and prevent its binding to the promoter. As reported earlier, DNA binding of the NF-κB subunit RelA is regulated by a variety of cofactors, such as ribosomal subunit S3, nucleoplasmin-1 and Sam68, among others (47)(48)(49)(50)(51), that directly bind to the Rel homology region (RHR) of RelA. While these cofactors support DNA binding of RelA, retinoblastoma protein (pRB), on the other hand, binds to the RHR of RelA and blocks it from binding to the promoters of some genes, such as Programmed death-ligand-1 (PD-L1) (52).…”
Section: Discussionmentioning
confidence: 74%
“…By 60 min, we also observed reduced levels of pSer142 on DPF2 (Zinc finger protein ubi-d4), a known regulator of the non-canonical NF-κB pathway [ 63 ], and pThr220/221 (site ambiguous) on the RelA binding partner RPS3 (40S ribosomal protein S3), both of which have previously been reported to be elevated in HEK293 cells following 3 h UV exposure [ 52 ]. Although binding of RSP3 to RelA is thought to enhance its ability to bind DNA in vitro [ 64 ], RSP3 functions as a negative regulator of H 2 O 2 -mediated DNA repair [ 65 ]. It is possible therefore that pThr220/221 on RPS3, and pSer142 on DPF2, serve to modulate the RelA transcriptional complexes and/or the consensus promoter regions for NF-κB binding, that are required for DNA repair in response to HU.…”
Section: Resultsmentioning
confidence: 99%
“…Consensus sequence motifs were generated with iceLogo v1.2 [ 43 ] against a background of the precompiled human Swiss-Prot composition using percent difference as the scoring system and a P -value cut-off of 0.05. The most confident phosphosite per phosphopeptide was also cross-referenced against data from PeptideAtlas (PA) [ 64 ] (2020 build), and from PhosphoSitePlus (PSP) [ 65 ] (11/03/20 build), categorising phosphorylation site confidence based on the number of observations - ‘High': ≥5 previous observations, very likely true site; ‘Medium': 2–4 previous observations, likely true site; ‘Low': 1 previous observation, little support that it is a true site; PA only – ‘Not phosphorylated': frequently (>5) observed to be not phosphorylated, never observed as phosphorylated; ‘Other' – no confident evidence in any category. Observations in PA were counted with a threshold of >0.95 PTM Prophet probability for positive evidence, and ≤0.19 for evidence of not being phosphorylated.…”
Section: Methodsmentioning
confidence: 99%