Dynamic changes in transcription factor complexes during erythroid differentiation revealed by quantitative proteomics

Brand, Marjorie; Ranish, Jeffrey A.; Kummer, Nicolas T; Hamilton, Joan; Igarashi, Kazuhiko; Francastel, Claire; Tian, Hui; Crabtree, Gerald R.; Aebersold, Ruedi; Groudine, Mark

doi:10.1038/nsmb713

Cited by 194 publications

(179 citation statements)

References 69 publications

Supporting

Mentioning

165

Contrasting

Unclassified

Order By: Relevance

“…It is possible that there may exist such an analogous anchor(s) for EKLF in the erythroid cell. Of more potential relevance to red cell biology, heme has been shown to affect the nuclear-cytoplasmic shuttling of Bach1 [49], a heterodimer partner of MafK that is known to be replaced by the critical p45/ NFE2 activator upon erythroid differentiation [50]. However, inhibition of heme synthesis by succinylacetone had no effect upon EKLF localization (unpublished observations).…”

Section: Discussionmentioning

confidence: 92%

Non-random subcellular distribution of variant EKLF in erythroid cells

Quadrini¹,

Gruzglin²,

Bieker³

2008

Experimental Cell Research

View full text Add to dashboard Cite

EKLF protein plays a prominent role during erythroid development as a nuclear transcription factor. Not surprisingly, exogenous EKLF quickly localizes to the nucleus. However, using two different assays we have unexpectedly found that a substantial proportion of endogenous EKLF resides in the cytoplasm at steady state in all erythroid cells examined. While EKLF localization does not appear to change during either erythroid development or terminal differentiation, we find that the protein displays subtle yet distinct biochemical and functional differences depending on which subcellular compartment it is isolated from, with PEST sequences possibly playing a role in these differences. Localization is unaffected by inhibition of CRM1 activity and the two populations are not differentiated by stability. Heterokaryon assays demonstrate that EKLF is able to shuttle out of the nucleus although its nuclear re-entry is rapid. These studies suggest there is an unexplored role for EKLF in the cytoplasm that is separate from its well-characterized nuclear function.

show abstract

Section: Discussionmentioning

confidence: 92%

Non-random subcellular distribution of variant EKLF in erythroid cells

Quadrini¹,

Gruzglin²,

Bieker³

2008

Experimental Cell Research

View full text Add to dashboard Cite

show abstract

“…The study of the cytoplasmic fraction of erythrocytes allowed the identification of many proteins, including hemoglobin a, b, and g chains. Quantitative proteomics was used by Brand et al [110] to evaluate the dynamic changes in transcription factor complexes during erythroid differentiation. The analysis of phosphoprotein profiling of erythropoietin receptor-dependent pathways was evaluated by Körbel et al [111], by combining two proteomic approaches (2-DE/MALDI-TOF; 1-D/LC-MS/MS).…”

Section: Red Blood Cellsmentioning

confidence: 99%

Recent advances in blood‐related proteomics

et al. 2005

View full text Add to dashboard Cite

Service régional vaudois de transfusion sanguine, Lausanne, SwitzerlandBlood is divided in two compartments, namely, plasma and cells. The latter contain red blood cells, leukocytes, and platelets. From a descriptive medical discipline, hematology has evolved towards a pioneering discipline where molecular biology has permitted the development of prognostic and diagnostic indicators for disease. The recent advance in MS and protein separation now allows similar progress in the analysis of proteins. Proteomics offers great promise for the study of proteins in plasma/serum, indeed a number of proteomics databases for plasma/ serum have been established. This is a very complex body fluid containing lipids, carbohydrates, amino acids, vitamins, nucleic acids, hormones, and proteins. About 1500 different proteins have recently been identified, and a number of potential new markers of diseases have been characterized. Here, examples of the enormous promise of plasma/serum proteomic analysis for diagnostic/prognostic markers and information on disease mechanism are given. Within the blood are also a large number of different blood cell types that potentially hold similar information. Proteomics of red blood cells, until now, has not improved our knowledge of these cells, in contrast to the major progresses achieved while studying platelets and leukocytes. In the future, proteomics will change several aspects of hematology.

show abstract

“…Protein-centered approaches aiming to characterize TF-specific protein interaction partners or complexes are also benefiting greatly from the recent sensitivity increase as evidenced by the fact that many of the detected TF interactors were themselves TFs. 37,38 Thus, we are entering an exciting era in which proteins such as TFs that have traditionally been for the most part off-limit become increasingly accessible and thus characterizable.…”

Section: Mass Spectrometrymentioning

confidence: 99%

“…The latter involve the labelling of proteins with isotopically distinguishable tags enabling a protein abundance comparison between two or more biological samples. Brand et al 37 used isotope-coded affinity tagging (ICAT) to monitor the compositional changes of the protein complex involving the TF NF-E2p18/MafK during erythroid differentiation. Results uncovered more than 100 potential protein interactors and indicated that MafK acts as a dual-function TF, exchanging dimerization partners upon induction of differentiation, leading to the replacement of interacting co-repressors with co-activators and up-regulation of the expression of its target gene b-globin.…”

Section: Dna-binding Dynamicsmentioning

confidence: 99%

DNA-centered approaches to characterize regulatory protein–DNA interaction complexes

Simicevic

Deplancke

2010

Mol. BioSyst.

View full text Add to dashboard Cite

Gene regulation is mediated by site-specific DNA-binding proteins or transcription factors (TFs), which form protein complexes at regulatory loci either to activate or repress the expression of a target gene. The study of the dynamic properties of these regulatory DNA-binding complexes has so far been dominated by protein-centered methodologies, aiming to characterize the DNA-binding behavior of one specific protein at a time. With the emerging evidence for a role of DNA in allosterically influencing DNA-binding protein complex formation, there is renewed interest in DNA-centered approaches to capture protein complexes on defined regulatory loci and to correlate changes in their composition with alterations in target gene expression. In this review, we present the current state-of-the-art in such DNA-centered approaches and evaluate recent technological improvements in the purification as well as in the identification of regulatory DNA-binding protein complexes within or outside their biological context. Finally, we suggest possible areas of improvement and assess the putative impact of DNA-centered methodologies on the gene regulation field for the forthcoming years.

show abstract

Dynamic changes in transcription factor complexes during erythroid differentiation revealed by quantitative proteomics

Cited by 194 publications

References 69 publications

Non-random subcellular distribution of variant EKLF in erythroid cells

Non-random subcellular distribution of variant EKLF in erythroid cells

Recent advances in blood‐related proteomics

DNA-centered approaches to characterize regulatory protein–DNA interaction complexes

Contact Info

Product

Resources

About