+Open data
-Basic information
Entry | Database: PDB / ID: 8pki | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Cryo-EM structure of NR5A2-nucleosome complex SHL+5.5 | |||||||||
Components |
| |||||||||
Keywords | DNA BINDING PROTEIN / Nucleosome / nuclear receptor / NR5A2 | |||||||||
Function / homology | Function and homology information Deposition of new CENPA-containing nucleosomes at the centromere / Inhibition of DNA recombination at telomere / SUMOylation of chromatin organization proteins / E3 ubiquitin ligases ubiquitinate target proteins / Regulation of gene expression in early pancreatic precursor cells / DNA Damage/Telomere Stress Induced Senescence / G2/M DNA damage checkpoint / HDMs demethylate histones / Regulation of endogenous retroelements by KRAB-ZFP proteins / Recognition and association of DNA glycosylase with site containing an affected purine ...Deposition of new CENPA-containing nucleosomes at the centromere / Inhibition of DNA recombination at telomere / SUMOylation of chromatin organization proteins / E3 ubiquitin ligases ubiquitinate target proteins / Regulation of gene expression in early pancreatic precursor cells / DNA Damage/Telomere Stress Induced Senescence / G2/M DNA damage checkpoint / HDMs demethylate histones / Regulation of endogenous retroelements by KRAB-ZFP proteins / Recognition and association of DNA glycosylase with site containing an affected purine / Condensation of Prophase Chromosomes / Cleavage of the damaged purine / Nonhomologous End-Joining (NHEJ) / pancreas morphogenesis / HDACs deacetylate histones / PRC2 methylates histones and DNA / Processing of DNA double-strand break ends / HATs acetylate histones / calcineurin-mediated signaling / PKMTs methylate histone lysines / Recruitment and ATM-mediated phosphorylation of repair and signaling proteins at DNA double strand breaks / RUNX1 regulates genes involved in megakaryocyte differentiation and platelet function / acinar cell differentiation / RMTs methylate histone arginines / tissue development / negative regulation of chromosome condensation / Barr body / regulation of centromere complex assembly / Factors involved in megakaryocyte development and platelet production / Estrogen-dependent gene expression / pericentric heterochromatin formation / inner kinetochore / bile acid metabolic process / muscle cell differentiation / embryo development ending in birth or egg hatching / oocyte maturation / oogenesis / homeostatic process / nucleus organization / chromosome, centromeric region / spermatid development / subtelomeric heterochromatin formation / single fertilization / RNA polymerase II core promoter sequence-specific DNA binding / positive regulation of viral genome replication / protein localization to CENP-A containing chromatin / CENP-A containing nucleosome / nucleosomal DNA binding / embryo implantation / hormone-mediated signaling pathway / cholesterol homeostasis / cellular response to leukemia inhibitory factor / transcription coregulator binding / SUMOylation of intracellular receptors / multicellular organism growth / phospholipid binding / Nuclear Receptor transcription pathway / osteoblast differentiation / RNA polymerase II transcription regulator complex / male gonad development / structural constituent of chromatin / nuclear receptor activity / sequence-specific double-stranded DNA binding / nucleosome / nucleosome assembly / chromatin organization / chromosome / regulation of cell population proliferation / spermatogenesis / positive regulation of cell growth / DNA-binding transcription activator activity, RNA polymerase II-specific / Estrogen-dependent gene expression / sequence-specific DNA binding / cell population proliferation / chromosome, telomeric region / transcription cis-regulatory region binding / DNA-binding transcription factor activity, RNA polymerase II-specific / protein heterodimerization activity / DNA-binding transcription factor activity / RNA polymerase II cis-regulatory region sequence-specific DNA binding / chromatin binding / regulation of DNA-templated transcription / chromatin / regulation of transcription by RNA polymerase II / positive regulation of DNA-templated transcription / positive regulation of transcription by RNA polymerase II / DNA binding / zinc ion binding / nucleoplasm / nucleus / cytoplasm Similarity search - Function | |||||||||
Biological species | Mus musculus (house mouse) synthetic construct (others) Homo sapiens (human) | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.58 Å | |||||||||
Authors | Kobayashi, W. / Sappler, A. / Bollschweiler, D. / Kummecke, M. / Basquin, J. / Arslantas, E. / Ruangroengkulrith, S. / Hornberger, R. / Duderstadt, K. / Tachibana, K. | |||||||||
Funding support | Germany, European Union, 2items
| |||||||||
Citation | Journal: Nat Struct Mol Biol / Year: 2024 Title: Nucleosome-bound NR5A2 structure reveals pioneer factor mechanism by DNA minor groove anchor competition. Authors: Wataru Kobayashi / Anna H Sappler / Daniel Bollschweiler / Maximilian Kümmecke / Jérôme Basquin / Eda Nur Arslantas / Siwat Ruangroengkulrith / Renate Hornberger / Karl Duderstadt / Kikuë Tachibana / Abstract: Gene expression during natural and induced reprogramming is controlled by pioneer transcription factors that initiate transcription from closed chromatin. Nr5a2 is a key pioneer factor that regulates ...Gene expression during natural and induced reprogramming is controlled by pioneer transcription factors that initiate transcription from closed chromatin. Nr5a2 is a key pioneer factor that regulates zygotic genome activation in totipotent embryos, pluripotency in embryonic stem cells and metabolism in adult tissues, but the mechanism of its pioneer activity remains poorly understood. Here, we present a cryo-electron microscopy structure of human NR5A2 bound to a nucleosome. The structure shows that the conserved carboxy-terminal extension (CTE) loop of the NR5A2 DNA-binding domain competes with a DNA minor groove anchor of the nucleosome and releases entry-exit site DNA. Mutational analysis showed that NR5A2 D159 of the CTE is dispensable for DNA binding but required for stable nucleosome association and persistent DNA 'unwrapping'. These findings suggest that NR5A2 belongs to an emerging class of pioneer factors that can use DNA minor groove anchor competition to destabilize nucleosomes and facilitate gene expression during reprogramming. | |||||||||
History |
|
-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
---|
-Downloads & links
-Download
PDBx/mmCIF format | 8pki.cif.gz | 390.4 KB | Display | PDBx/mmCIF format |
---|---|---|---|---|
PDB format | pdb8pki.ent.gz | 239.8 KB | Display | PDB format |
PDBx/mmJSON format | 8pki.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8pki_validation.pdf.gz | 1.4 MB | Display | wwPDB validaton report |
---|---|---|---|---|
Full document | 8pki_full_validation.pdf.gz | 1.4 MB | Display | |
Data in XML | 8pki_validation.xml.gz | 43 KB | Display | |
Data in CIF | 8pki_validation.cif.gz | 65.6 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/pk/8pki ftp://data.pdbj.org/pub/pdb/validation_reports/pk/8pki | HTTPS FTP |
-Related structure data
Related structure data | 17740MC 8pkjC M: map data used to model this data C: citing same article (ref.) |
---|---|
Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
|
---|---|
1 |
|
-Components
-Protein , 5 types, 9 molecules AEBFCGDHK
#1: Protein | Mass: 15360.983 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mus musculus (house mouse) / Gene: H3-3a, H3.3a, H3f3a, H3-3b, H3.3b, H3f3b / Production host: Escherichia coli (E. coli) / References: UniProt: P84244 #2: Protein | Mass: 11394.426 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mus musculus (house mouse) Gene: H4c1, Hist1h4a, H4c2, H4-53, Hist1h4b, H4c3, H4-12, Hist1h4c, H4c4, Hist1h4d, H4c6, Hist1h4f, H4c8, Hist1h4h, H4c9, Hist1h4i, H4c11, Hist1h4j, H4c12, Hist1h4k, Hist1h4m, H4c14, Hist2h4, ...Gene: H4c1, Hist1h4a, H4c2, H4-53, Hist1h4b, H4c3, H4-12, Hist1h4c, H4c4, Hist1h4d, H4c6, Hist1h4f, H4c8, Hist1h4h, H4c9, Hist1h4i, H4c11, Hist1h4j, H4c12, Hist1h4k, Hist1h4m, H4c14, Hist2h4, Hist2h4a, H4c16, H4f16, Hist4h4 Production host: Escherichia coli (E. coli) / References: UniProt: P62806 #3: Protein | Mass: 14165.551 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mus musculus (house mouse) / Gene: Hist1h2ad / Production host: Escherichia coli (E. coli) / References: UniProt: B2RVF0 #4: Protein | Mass: 13937.213 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mus musculus (house mouse) / Gene: H2bc4, Hist1h2bc, H2bc6, Hist1h2be, H2bc8, Hist1h2bg / Production host: Escherichia coli (E. coli) / References: UniProt: Q6ZWY9 #7: Protein | | Mass: 10989.939 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: NR5A2, B1F, CPF, FTF / Production host: Escherichia coli (E. coli) / References: UniProt: O00482 |
---|
-DNA chain , 2 types, 2 molecules IJ
#5: DNA chain | Mass: 46968.922 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) synthetic construct (others) / Production host: Escherichia coli (E. coli) |
---|---|
#6: DNA chain | Mass: 47489.234 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) synthetic construct (others) / Production host: Escherichia coli (E. coli) |
-Non-polymers , 1 types, 1 molecules
#8: Chemical | ChemComp-ZN / |
---|
-Details
Has ligand of interest | Y |
---|
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
---|---|
EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Cryo-EM structure of the nucleosome containing NR5A2 motif at SHL+5.5 Type: COMPLEX / Entity ID: #1-#7 / Source: RECOMBINANT |
---|---|
Molecular weight | Experimental value: NO |
Source (natural) | Organism: Mus musculus (house mouse) |
Source (recombinant) | Organism: Escherichia coli (E. coli) |
Buffer solution | pH: 7.5 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
---|---|
Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: SPOT SCAN |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2200 nm / Nominal defocus min: 600 nm |
Image recording | Electron dose: 65.4 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
3D reconstruction | Resolution: 2.58 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 653440 / Symmetry type: POINT | ||||||||||||||||||||||||
Refinement | Cross valid method: NONE Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2 | ||||||||||||||||||||||||
Displacement parameters | Biso mean: 110.27 Å2 | ||||||||||||||||||||||||
Refine LS restraints |
|