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Title: Double DAP-seq uncovered synergistic DNA binding of interacting bZIP transcription factors

Journal Article · · Nature Communications

Many eukaryotic transcription factors (TF) form homodimer or heterodimer complexes to regulate gene expression. Dimerization of BASIC LEUCINE ZIPPER (bZIP) TFs are critical for their functions, but the molecular mechanism underlying the DNA binding and functional specificity of homo- versus heterodimers remains elusive. To address this gap, we present the double DNA Affinity Purification-sequencing (dDAP-seq) technique that maps heterodimer binding sites on endogenous genomic DNA. Using dDAP-seq we profile twenty pairs of C/S1 bZIP heterodimers and S1 homodimers in Arabidopsis and show that heterodimerization significantly expands the DNA binding preferences of these TFs. Analysis of dDAP-seq binding sites reveals the function of bZIP9 in abscisic acid response and the role of bZIP53 heterodimer-specific binding in seed maturation. The C/S1 heterodimers show distinct preferences for the ACGT elements recognized by plant bZIPs and motifs resembling the yeast GCN4 cis-elements. This study demonstrates the potential of dDAP-seq in deciphering the DNA binding specificities of interacting TFs that are key for combinatorial gene regulation.

Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); National Science Foundation (NSF)
Grant/Contract Number:
AC05-00OR22725; R35GM138143; 1T32GM132037-01; IOS-1916804
OSTI ID:
1994639
Journal Information:
Nature Communications, Vol. 14, Issue 1; ISSN 2041-1723
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
English

References (122)

Combinatorial Regulation of Endothelial Gene Expression by Ets and Forkhead Transcription Factors journal December 2008
Genome-Wide Characterization ofcis-Acting DNA Targets Reveals the Transcriptional Regulatory Framework ofOpaque2in Maize journal February 2015
Two bZIP proteins fromAntirrhinumflowers preferentially bind a hybrid C‐box/G‐box motif and help to define a new sub‐family of bZIP transcription factors journal February 1998
Model-based Analysis of ChIP-Seq (MACS) journal January 2008
MEME-ChIP: motif analysis of large DNA datasets journal April 2011
SMiLE-seq identifies binding motifs of single and dimeric transcription factors journal January 2017
Direct Control of SPEECHLESS by PIF4 in the High-Temperature Response of Stomatal Development journal April 2018
Impact of artifact removal on ChIP quality metrics in ChIP-seq and ChIP-exo data journal April 2014
MADS transcription factors cooperate: complexities of complex formation journal April 2018
Transcriptional program for nitrogen starvation-induced lipid accumulation in Chlamydomonas reinhardtii journal December 2015
Heterodimers of the Arabidopsis Transcription Factors bZIP1 and bZIP53 Reprogram Amino Acid Metabolism during Low Energy Stress journal January 2011
Determination and Inference of Eukaryotic Transcription Factor Sequence Specificity journal September 2014
Abscisic Acid: Emergence of a Core Signaling Network journal June 2010
The role of phosphorylatable serine residues in the DNA-binding domain of Arabidopsis bZIP transcription factors journal February 2010
Discovering Motifs in Ranked Lists of DNA Sequences journal March 2007
Barley BLZ2, a Seed-specific bZIP Protein That Interacts with BLZ1 in Vivo and Activates Transcription from the GCN4-like motif of B-hordein Promoters in Barley Endosperm journal April 1999
The Chlamydomonas bZIP transcription factor BLZ8 confers oxidative stress tolerance by inducing the carbon-concentrating mechanism journal December 2021
Synergistic Activation of Seed Storage Protein Gene Expression in Arabidopsis by ABI3 and Two bZIPs Related to OPAQUE2 journal March 2003
Comparison of discriminative motif optimization using matrix and DNA shape-based models journal March 2018
DNA-dependent formation of transcription factor pairs alters their binding specificity journal November 2015
MAnorm2 for quantitatively comparing groups of ChIP-seq samples journal November 2020
Context of action of Proline Dehydrogenase (ProDH) in the Hypersensitive Response of Arabidopsis journal January 2014
The AtGenExpress hormone and chemical treatment data set: experimental design, data evaluation, model data analysis and data access journal August 2008
Expression patterns within the Arabidopsis C/S1 bZIP transcription factor network: availability of heterodimerization partners controls gene expression during stress response and development journal October 2008
Changing MADS-Box Transcription Factor Protein–Protein Interactions as a Mechanism for Generating Floral Morphological Diversity journal July 2017
A systematic genome-wide account of binding sites for the model transcription factor Gcn4 journal December 2021
Characterization of abiotic stress-responsive Arabidopsis thaliana RD29A and RD29B genes and evaluation of transgenes journal March 2011
Transcriptional activation by recruitment journal April 1997
bZIPs and WRKYs: two large transcription factor families executing two different functional strategies journal April 2014
Response of plant metabolism to too little oxygen journal June 2003
GCN4 Binds with High Affinity to DNA Sequences Containing a Single Consensus Half-Site journal May 2000
Mitochondrial signalling is critical for acclimation and adaptation to flooding in Arabidopsis thaliana journal April 2020
B-ZIP Proteins Encoded by the Drosophila Genome: Evaluation of Potential Dimerization Partners journal August 2002
The Roles of Auxin Response Factor Domains in Auxin-Responsive Transcription journal January 2003
A Pivotal Role of the Basic Leucine Zipper Transcription Factor bZIP53 in the Regulation of Arabidopsis Seed Maturation Gene Expression Based on Heterodimerization and Protein Complex Formation journal June 2009
Mass-spectrometry-based draft of the Arabidopsis proteome journal March 2020
deepTools2: a next generation web server for deep-sequencing data analysis journal April 2016
Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis journal June 2007
PlantPAN3.0: a new and updated resource for reconstructing transcriptional regulatory networks from ChIP-seq experiments in plants journal November 2018
ChIP–seq: advantages and challenges of a maturing technology journal September 2009
Arabidopsis Transcription Factors: Genome-Wide Comparative Analysis Among Eukaryotes journal December 2000
ACTCAT, a Novel cis-Acting Element for Proline- and Hypoosmolarity-Responsive Expression of the ProDH Gene Encoding Proline Dehydrogenase in Arabidopsis journal October 2002
bZIP transcription factors in Arabidopsis journal March 2002
Genome-wide identification of grain filling genes regulated by the OsSMF1 transcription factor in rice journal April 2017
A central integrator of transcription networks in plant stress and energy signalling journal August 2007
Genome-wide binding of SEPALLATA3 and AGAMOUS complexes determined by sequential DNA-affinity purification sequencing journal September 2020
A novel k-mer set memory (KSM) motif representation improves regulatory variant prediction journal April 2018
Plant bZIP proteins gather at ACGT elements journal February 1994
The Origin of Land Plants Is Rooted in Two Bursts of Genomic Novelty journal February 2020
Transcription factor–DNA binding: beyond binding site motifs journal April 2017
Homotypic cooperativity and collective binding are determinants of bHLH specificity and function journal July 2019
Plant bZIP Protein DNA Binding Specificity journal April 1993
Abscisic Acid Signaling in Seeds and Seedlings journal May 2002
Local DNA shape is a general principle of transcription factor binding specificity in Arabidopsis thaliana journal November 2021
GO-PCA: An Unsupervised Method to Explore Gene Expression Data Using Prior Knowledge journal November 2015
Non-targeted transcription factors motifs are a systemic component of ChIP-seq datasets journal July 2014
Differential analysis of RNA-seq incorporating quantification uncertainty journal June 2017
A Novel Subgroup of bZIP Proteins Functions as Transcriptional Activators in Hypoosmolarity-Responsive Expression of the ProDH Gene in Arabidopsis journal March 2004
OXYGEN DEFICIENCY AND ROOT METABOLISM: Injury and Acclimation Under Hypoxia and Anoxia journal June 1997
A Triple Helix-Loop-Helix/Basic Helix-Loop-Helix Cascade Controls Cell Elongation Downstream of Multiple Hormonal and Environmental Signaling Pathways in Arabidopsis journal December 2012
Mapping genome-wide transcription-factor binding sites using DAP-seq journal July 2017
Cistrome and Epicistrome Features Shape the Regulatory DNA Landscape journal May 2016
Assessing Computational Methods for Transcription Factor Target Gene Identification Based on ChIP-seq Data journal November 2013
Complex heatmaps reveal patterns and correlations in multidimensional genomic data journal May 2016
Diversity and specificity in transcriptional regulation: the benefits of heterotypic dimerization journal January 1991
Cutadapt removes adapter sequences from high-throughput sequencing reads journal May 2011
A Rice Functional Transcriptional Activator, RISBZ1, Responsible for Endosperm-specific Expression of Storage Protein Genes through GCN4 Motif journal April 2001
Plant transcription factors — being in the right place with the right company journal February 2022
DiffLogo: a comparative visualization of sequence motifs journal November 2015
Mapping and Dynamics of Regulatory DNA and Transcription Factor Networks in A. thaliana journal September 2014
The Arabidopsis bZIP transcription factor family — an update journal October 2018
Metabolic Regulation of the Gene Encoding Glutamine-Dependent Asparagine Synthetase in Arabidopsis thaliana journal December 1994
The G-Box Transcriptional Regulatory Code in Arabidopsis journal September 2017
A-ZIP53, a dominant negative reveals the molecular mechanism of heterodimerization between bZIP53, bZIP10 and bZIP25 involved in Arabidopsis seed maturation journal October 2017
The Arabidopsis transcription factor bZIP11 activates auxin-mediated transcription by recruiting the histone acetylation machinery journal May 2014
Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of Yeast journal September 1986
Curated collection of yeast transcription factor DNA binding specificity data reveals novel structural and gene regulatory insights journal January 2011
The GCN4 bZIP Targets Noncognate Gene Regulatory Sequences:  Quantitative Investigation of Binding at Full and Half Sites journal January 2007
The Regulatory Code for Transcriptional Response Diversity and Its Relation to Genome Structural Properties in A. thaliana journal February 2007
The GCN4 basic region leucine zipper binds DNA as a dimer of uninterrupted α Helices: Crystal structure of the protein-DNA complex journal December 1992
The Next Generation of Transcription Factor Binding Site Prediction journal September 2013
Identification and functional characterization of transcriptional activators in human cells journal February 2022
The C/S1 bZIP Network: A Regulatory Hub Orchestrating Plant Energy Homeostasis journal May 2018
The Arabidopsis bZIP1 Transcription Factor Is Involved in Sugar Signaling, Protein Networking, and DNA Binding journal March 2010
Varying levels of complexity in transcription factor binding motifs journal June 2015
Structural Basis for DNA Binding Specificity by the Auxin-Dependent ARF Transcription Factors journal January 2014
Snf1-RELATED KINASE1-Controlled C/S 1 -bZIP Signaling Activates Alternative Mitochondrial Metabolic Pathways to Ensure Plant Survival in Extended Darkness journal January 2018
In pursuit of design principles of regulatory sequences journal June 2014
bZIP10-LSD1 antagonism modulates basal defense and cell death in Arabidopsis following infection journal September 2006
One Billion Years of bZIP Transcription Factor Evolution: Conservation and Change in Dimerization and DNA-Binding Site Specificity journal December 2006
EnrichedHeatmap: an R/Bioconductor package for comprehensive visualization of genomic signal associations journal April 2018
The mammalian Jun proteins: redundancy and specificity journal April 2001
The Elucidation of the Interactome of 16 Arabidopsis bZIP Factors Reveals Three Independent Functional Networks journal October 2015
Dimerization specificity of all 67 B-ZIP motifs in Arabidopsis thaliana: a comparison to Homo sapiens B-ZIP motifs journal June 2004
The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins journal June 1988
ggseqlogo: a versatile R package for drawing sequence logos journal July 2017
karyoploteR: an R/Bioconductor package to plot customizable genomes displaying arbitrary data journal May 2017
One-Step Purification of Recombinant Proteins Using a Nanomolar-Affinity Streptavidin-Binding Peptide, the SBP-Tag journal December 2001
High-resolution DNA-binding specificity analysis of yeast transcription factors journal January 2009
Multiple protein factors bind to a rice glutelin promoter region journal January 1990
Low-Affinity Binding Sites and the Transcription Factor Specificity Paradox in Eukaryotes journal October 2019
The Arabidopsis NPR1/NIM1 Protein Enhances the DNA Binding Activity of a Subgroup of the TGA Family of bZIP Transcription Factors journal February 2000
The phylogeny of C/S1 bZIP transcription factors reveals a shared algal ancestry and the pre-angiosperm translational regulation of S1 transcripts journal July 2016
The oxygen status of the developing seed journal January 2009
The Role of bZIP Transcription Factors in Green Plant Evolution: Adaptive Features Emerging from Four Founder Genes journal August 2008
Transcription factor enrichment analysis (TFEA) quantifies the activity of multiple transcription factors from a single experiment journal June 2021
Combinatorial control of Arabidopsis proline dehydrogenase transcription by specific heterodimerisation of bZIP transcription factors journal June 2006
Fast gapped-read alignment with Bowtie 2 journal March 2012
Combinatorial Transcriptional Control of Plant Specialized Metabolism journal April 2018
Divergence in DNA Specificity among Paralogous Transcription Factors Contributes to Their Differential In Vivo Binding journal April 2018
Interaction of NPR1 with basic leucine zipper protein transcription factors that bind sequences required for salicylic acid induction of the PR-1 gene journal May 1999
High Resolution Genome Wide Binding Event Finding and Motif Discovery Reveals Transcription Factor Spatial Binding Constraints journal August 2012
Combinatorial bZIP dimers display complex DNA-binding specificity landscapes journal February 2017
DNA-binding specificities of plant transcription factors and their potential to define target genes journal January 2014
Universal protein-binding microarrays for the comprehensive characterization of the DNA-binding specificities of transcription factors journal March 2009
A Novel Wheat C-bZIP Gene, TabZIP14-B, Participates in Salt and Freezing Tolerance in Transgenic Plants journal May 2017
Two-hybrid protein-protein interaction analysis in Arabidopsis protoplasts: establishment of a heterodimerization map of group C and group S bZIP transcription factors journal June 2006
Adapterama I: universal stubs and primers for 384 unique dual-indexed or 147,456 combinatorially-indexed Illumina libraries (iTru & iNext) journal January 2019
Hit-and-run transcriptional control by bZIP1 mediates rapid nutrient signaling in Arabidopsis journal June 2014
Near-optimal probabilistic RNA-seq quantification journal April 2016
SnRK1-triggered switch of bZIP63 dimerization mediates the low-energy response in plants journal August 2015
A transcription factor hierarchy defines an environmental stress response network journal November 2016