Steric Mechanism of Auto-Inhibitory Regulation of Specific and Non-Specific DNA Binding by the ETS Transcriptional Repressor ETV6

De, Soumya; Chan, Anson; Coyne, H. Jerome; Bhachech, Niraja; Hermsdorf, Ulrike; Okon, Mark; Murphy, M.E.P.; Graves, Barbara J.; McIntosh, Lawrence P.

doi:10.1016/j.jmb.2013.11.031

Cited by 48 publications

(61 citation statements)

References 80 publications

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“…Furthermore, ERG lacks an equivalent IM and is not known to be modulated at the level of DNA binding by phosphorylation. In contrast, ETV6 does not contain an equivalent SRR and instead is autoinhibited by a C-terminal helix that sterically blocks its ETS domain interface and unfolds upon binding both specific and nonspecific DNA sequences (28). In all of these cases, additional DNA binding proteins can counteract or bypass the negative effects of autoinhibition and thereby provide added specificity to each ETS factor.…”

Section: Discussionmentioning

confidence: 99%

Synergy of aromatic residues and phosphoserines within the intrinsically disordered DNA-binding inhibitory elements of the Ets-1 transcription factor

Desjardins

Meeker

Bhachech

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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The E26 transformation-specific (Ets-1) transcription factor is autoinhibited by a conformationally disordered serine-rich region (SRR) that transiently interacts with its DNA-binding ETS domain. In response to calcium signaling, autoinhibition is reinforced by calmodulin-dependent kinase II phosphorylation of serines within the SRR. Using mutagenesis and quantitative DNA-binding measurements, we demonstrate that phosphorylation-enhanced autoinhibition requires the presence of phenylalanine or tyrosine (ϕ) residues adjacent to the SRR phosphoacceptor serines. The introduction of additional phosphorylated Ser-ϕ-Asp, but not Ser-Ala-Asp, repeats within the SRR dramatically reinforces autoinhibition. NMR spectroscopic studies of phosphorylated and mutated SRR variants, both within their native context and as separate trans-acting peptides, confirmed that the aromatic residues and phosphoserines contribute to the formation of a dynamic complex with the ETS domain. Complementary NMR studies also identified the SRR-interacting surface of the ETS domain, which encompasses its positively charged DNArecognition interface and an adjacent region of neutral polar and nonpolar residues. Collectively, these studies highlight the role of aromatic residues and their synergy with phosphoserines in an intrinsically disordered regulatory sequence that integrates cellular signaling and gene expression.transcription factor regulation | protein dynamics | intrinsically disordered region | fuzz complex I ntrinsically disordered protein regions (IDRs) are increasingly recognized for their prevalence in the eukaryotic proteome and their roles in normal biological processes, as well as in disease (1-3). IDRs serve as flexible linker sequences between modular domains and as key components of complex protein-interaction networks. These sequences are often sites of posttranslational modifications, and their plasticity enables accessible and reversible interactions necessary for the integration of cellular signals.The autoinhibition of the E26 transformation-specific (Ets-1) transcription factor provides an illuminating example of how a flexible, disordered region can modulate the regulatable DNAbinding ETS domain and thereby tune it for biological control by calcium-dependent phosphorylation and cooperative protein partnerships (4). The inhibitory module (IM) of Ets-1 is composed of four α-helices (HI-1, HI-2, H4, and H5) that pack onto the ETS domain distal from the DNA interface (Fig. 1A) (5). Helix HI-1 is marginally stable and unfolds on DNA binding, thus implicating an allosteric mechanism of inhibition (6, 7). The modest twofold repression afforded by the IM is increased to ∼20-fold by an intrinsically disordered serine-rich region (SRR) (8, 9). The dynamic SRR interacts transiently with both the ETS domain DNA-binding interface and the IM, and thus plays steric and allosteric inhibitory roles (10). Phosphorylation of five SRR serines by calmodulindependent kinase II (CaMKII) leads to a dramatic ∼500-fold autoinhibition (11). In paral...

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Section: Discussionmentioning

confidence: 99%

Synergy of aromatic residues and phosphoserines within the intrinsically disordered DNA-binding inhibitory elements of the Ets-1 transcription factor

Desjardins

Meeker

Bhachech

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…The Thermodynamics and Kinetics of Ets-1 AutoinhibitionThe structure and dynamics of autoinhibition in Ets-1 (35)(36)(37)(38)(39)(40), as well as other autoinhibited ETS paralogs (58,59), have been extensively studied by solution NMR. The folded helices immediately adjacent to the ETS domain of Ets-1 are only marginally stable in the unbound state and unfold upon DNA binding.…”

Section: Discussionmentioning

confidence: 99%

Mechanistic Heterogeneity in Site Recognition by the Structurally Homologous DNA-binding Domains of the ETS Family Transcription Factors Ets-1 and PU.1

Wang

Linde

Munde

et al. 2014

Journal of Biological Chemistry

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Background: ETS family transcription factors recognize DNA via structurally conserved DNA-binding domains that share limited amino acid homology. Results: DNA recognition by the ETS domains of Ets-1 and PU.1, two extreme sequence-divergent paralogs, was compared. Conclusion: Preferential hydration differentiates DNA recognition by Ets-1 and PU.1. Significance: Preferential hydration represents a potential mechanism for PU.1 regulation and its activity as a pioneer transcription factor in vivo.

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“…12 A second ETS member, ETV6, a transcriptional repressor, 13 has also been found recently to be autoinhibited by Helix H5 in C inhibitory domain, which would unfold when binding to DNA. 14,15 DNA-binding autoinhibition has also been reported for other ETS factors. For example, members of the PEA3 subfamily have inhibitory sequences, but do not appear to be helical as determined by proline-scanning mutagenesis.…”

Section: Introductionmentioning

confidence: 68%

Autoinhibitory mechanisms of ERG studied by molecular dynamics simulations

Lü

Salsbury

2015

AIP Advances

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ERG, an ETS-family transcription factor, acts as a regulator of differentiation of early hematopoietic cells. It contains an autoinhibitory domain, which negatively regulates DNA-binding. The mechanism of autoinhibitory is still illusive. To understand the mechanism, we study the dynamical properties of ERG protein by molecular dynamics simulations. These simulations suggest that DNA binding autoinhibition associates with the internal dynamics of ERG. Specifically, we find that (1), The N-C terminal correlation in the inhibited ERG is larger than that in uninhibited ERG that contributes to the autoinhibition of DNA-binding. (2), DNA-binding changes the property of the N-C terminal correlation from being anti-correlated to correlated, that is, changing the relative direction of the correlated motions and (3), For the Ets-domain specifically, the inhibited and uninhibited forms exhibit essentially the same dynamics, but the binding of the DNA decreases the fluctuation of the Ets-domain. We also find from PCA analysis that the three systems, even with quite different dynamics, do have highly similar free energy surfaces, indicating that they share similar conformations.

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Steric Mechanism of Auto-Inhibitory Regulation of Specific and Non-Specific DNA Binding by the ETS Transcriptional Repressor ETV6

Cited by 48 publications

References 80 publications

Synergy of aromatic residues and phosphoserines within the intrinsically disordered DNA-binding inhibitory elements of the Ets-1 transcription factor

Synergy of aromatic residues and phosphoserines within the intrinsically disordered DNA-binding inhibitory elements of the Ets-1 transcription factor

Mechanistic Heterogeneity in Site Recognition by the Structurally Homologous DNA-binding Domains of the ETS Family Transcription Factors Ets-1 and PU.1

Autoinhibitory mechanisms of ERG studied by molecular dynamics simulations

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