Quantifying the dynamics of the oligomeric transcription factor STAT3 by pair correlation of molecular brightness

Hinde, Elizabeth; Pandžić, Elvis; Yang, Zhengmin; Ng, Ivan Hong Wee; Jans, David A.; Bogoyevitch, Marie A.; Gratton, Enrico; Gaus, Katharina

doi:10.1038/ncomms11047

Cited by 31 publications

(49 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Early biochemical studies showed the formation of STAT tetramers on repeated motifs with a spacing of 11-12 bp (Vinkemeier et al 1998). Recent evidence supporting the conversion of STAT3 from dimers to tetramers during DNA binding in living cells was presented by Gaus and colleagues (Hinde et al 2016), using the pCOMB technique.…”

Section: Discussionmentioning

confidence: 99%

“…The bound TP53 protein (also known as p53) has been described as a tetramer, based on both static methods (Emamzadah et al 2011;Rhee and Pugh 2011) and live cell approaches (Gaglia et al 2013). More recently, the DNA-bound STAT3 factor was also confirmed to be a tetramer, based on pair correlation of molecular brightness (pCOMB), another live cell technique (Hinde et al 2016). These findings illustrate the inadequacy of static cell data sets for an accurate and complete understanding of molecular status for regulatory complexes.…”

mentioning

confidence: 98%

See 1 more Smart Citation

Glucocorticoid receptor quaternary structure drives chromatin occupancy and transcriptional outcome

et al. 2019

View full text Add to dashboard Cite

Most transcription factors, including nuclear receptors, are widely modeled as binding regulatory elements as monomers, homodimers, or heterodimers. Recent findings in live cells show that the glucocorticoid receptor NR3C1 (also known as GR) forms tetramers on enhancers, owing to an allosteric alteration induced by DNA binding, and suggest that higher oligomerization states are important for the gene regulatory responses of GR. By using a variant (GRtetra) that mimics this allosteric transition, we performed genome-wide studies using a GR knockout cell line with reintroduced wild-type GR or reintroduced GRtetra. GRtetra acts as a super receptor by binding to response elements not accessible to the wild-type receptor and both induces and represses more genes than GRwt. These results argue that DNA binding induces a structural transition to the tetrameric state, forming a transient higher-order structure that drives both the activating and repressive actions of glucocorticoids.[Supplemental material is available for this article.] 4 These authors contributed equally to this work. Corresponding authors: hagerg@dce41.nci.nih.gov, presmandm@fbmc.fcen.uba.ar Article published online before print. Article, supplemental material, and publication date are at http://www.genome.org/cgi/

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 98%

Glucocorticoid receptor quaternary structure drives chromatin occupancy and transcriptional outcome

et al. 2019

View full text Add to dashboard Cite

show abstract

“…DNA-dependent tetramerization of transcription factors may be a more common phenomenon than originally thought. For example, a recent publication has shown that the signal transducer and activator of transcription 3 (STAT3) dimers must first bind DNA to form STAT3 tetramers (36). A reevaluation of several known dimeric transcription factors with these new state-of-the-art techniques might bring some surprises to the field.…”

Section: Respectively) (E)mentioning

confidence: 99%

DNA binding triggers tetramerization of the glucocorticoid receptor in live cells

Presman

Ganguly

et al. 2016

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

119

190

View full text Add to dashboard Cite

Transcription factors dynamically bind to chromatin and are essential for the regulation of genes. Although a large percentage of these proteins appear to self-associate to form dimers or higher order oligomers, the stoichiometry of DNA-bound transcription factors has been poorly characterized in vivo. The glucocorticoid receptor (GR) is a ligandregulated transcription factor widely believed to act as a dimer or a monomer. Using a unique set of imaging techniques coupled with a cell line containing an array of DNA binding elements, we show that GR is predominantly a tetramer when bound to its target DNA. We find that DNA binding triggers an interdomain allosteric regulation within the GR, leading to tetramerization. We therefore propose that dynamic changes in GR stoichiometry represent a previously unidentified level of regulation in steroid receptor activation. Quaternary structure analysis of other members of the steroid receptor family (estrogen, androgen, and progesterone receptors) reveals variation in oligomerization states among this family of transcription factors. Because GR's oligomerization state has been implicated in therapy outcome, our findings open new doors to the rational design of novel GR ligands and redefine the quaternary structure of steroid receptors. Steroid receptors are transcription factors regulated by physiological stimuli that dynamically bind to chromatin and control complex biological pathways (1). In particular, the glucocorticoid receptor (GR) is essential for life and is one of the most targeted proteins in the pharmacological industry due to its powerful antiinflammatory and immunosuppressive activities (2). Current pharmaceutical approaches are based on a recently challenged (3) binary model wherein direct binding of GR dimers and indirect binding of GR monomers via other proteins determine the transcriptional output (4).Upon hormone activation, GR associates to a subset of glucocorticoid response elements (GREs) across the genome, depending on the accessibility of the chromatin landscape (5). GR is a modular protein organized into three structural and functional domains; the N-terminal ligand-independent activation function-1 domain (NTD), the central DNA-binding domain (DBD), and the C-terminal ligand-binding domain (LBD) (6). GR, and all steroid receptors, are widely believed to bind DNA directly as homodimers (7). However, this paradigm has been established exclusively from in vitro studies, working mostly with the DBD fragment (8-10), only using the whole GR protein in rare cases (11,12). The small number of experiments performed in live cells only addresses the entire nuclear population, lacking specific information regarding the GR fraction bound to chromatin (13-16). Furthermore, these studies were unable to discriminate between dimers or higher oligomeric states.For the present study, we combine an experimental model where GR-DNA interaction can be observed in real time with techniques that allow the quantification of the oligomeric state of proteins inside living...

show abstract

“…For example, FCS measurements of p53 in single cells reveal that the TF exists in a mixture of oligomeric states; their rapid homo-tetramerization upon DNA damage activates the transcriptional targets of p53 [121]. Similar strategies have also been observed for the activation of the Rac1 signaling pathway [122], interaction between HP1-α and heterochromatin [123], and DNA-binding of STAT3 upon nuclear translocation [124] (Fig. 4b).…”

Section: Regulating Transcriptional Dynamics Through Concentration Omentioning

confidence: 84%

“…c Immunofluorescence staining of four-cell mouse embryos reveals distinct differences in H3R26me2 levels ( top ); cells with a higher level ( orange ) exhibit a larger fraction of Sox2 engaged in long-lived DNA-binding than those with a lower level ( blue ) ( bottom ). Adapted from [118] ( a ), [124] ( b ), and [66] ( c ) with modifications …”

Section: Regulating Transcriptional Dynamics Through Concentration Omentioning

confidence: 99%

Quantitative imaging of mammalian transcriptional dynamics: from single cells to whole embryos

et al. 2016

View full text Add to dashboard Cite

Probing dynamic processes occurring within the cell nucleus at the quantitative level has long been a challenge in mammalian biology. Advances in bio-imaging techniques over the past decade have enabled us to directly visualize nuclear processes in situ with unprecedented spatial and temporal resolution and single-molecule sensitivity. Here, using transcription as our primary focus, we survey recent imaging studies that specifically emphasize the quantitative understanding of nuclear dynamics in both time and space. These analyses not only inform on previously hidden physical parameters and mechanistic details, but also reveal a hierarchical organizational landscape for coordinating a wide range of transcriptional processes shared by mammalian systems of varying complexity, from single cells to whole embryos.

show abstract

Quantifying the dynamics of the oligomeric transcription factor STAT3 by pair correlation of molecular brightness

Cited by 31 publications

References 54 publications

Glucocorticoid receptor quaternary structure drives chromatin occupancy and transcriptional outcome

Glucocorticoid receptor quaternary structure drives chromatin occupancy and transcriptional outcome

DNA binding triggers tetramerization of the glucocorticoid receptor in live cells

Quantitative imaging of mammalian transcriptional dynamics: from single cells to whole embryos

Contact Info

Product

Resources

About