U2 snRNA 3'-end processing / NELF complex / meiotic spindle elongation / Integration of energy metabolism / NTRK3 as a dependence receptor / PP2A-mediated dephosphorylation of key metabolic factors / snRNA 3'-end processing / mitotic sister chromatid separation / MASTL Facilitates Mitotic Progression / negative regulation of DNA-templated transcription, elongation ...U2 snRNA 3'-end processing / NELF complex / meiotic spindle elongation / Integration of energy metabolism / NTRK3 as a dependence receptor / PP2A-mediated dephosphorylation of key metabolic factors / snRNA 3'-end processing / mitotic sister chromatid separation / MASTL Facilitates Mitotic Progression / negative regulation of DNA-templated transcription, elongation / protein phosphatase type 2A complex / RNA polymerase II CTD heptapeptide repeat S2 phosphatase activity / RNA polymerase II CTD heptapeptide repeat S7 phosphatase activity / protein serine/threonine phosphatase complex / regulation of meiotic cell cycle process involved in oocyte maturation / peptidyl-threonine dephosphorylation / meiotic sister chromatid cohesion, centromeric / snRNA processing / : / INTAC complex / FAR/SIN/STRIPAK complex / RNA polymerase II CTD heptapeptide repeat S5 phosphatase activity / Regulation of glycolysis by fructose 2,6-bisphosphate metabolism / DSIF complex / Inhibition of replication initiation of damaged DNA by RB1/E2F1 / female meiotic nuclear division / regulation of transcription elongation by RNA polymerase II / protein phosphatase regulator activity / GABA receptor binding / integrator complex / APC truncation mutants have impaired AXIN binding / AXIN missense mutants destabilize the destruction complex / Truncations of AMER1 destabilize the destruction complex / protein antigen binding / Formation of RNA Pol II elongation complex / Formation of the Early Elongation Complex / Transcriptional regulation by small RNAs / RNA Polymerase II Pre-transcription Events / TP53 Regulates Transcription of DNA Repair Genes / FGFR2 alternative splicing / RNA polymerase II transcribes snRNA genes / mRNA Capping / mRNA Splicing - Minor Pathway / Processing of Capped Intron-Containing Pre-mRNA / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening / RNA Polymerase II Transcription Initiation / RNA Polymerase II Transcription Elongation / RNA Polymerase II Transcription Initiation And Promoter Clearance / RNA Pol II CTD phosphorylation and interaction with CE / Estrogen-dependent gene expression / Formation of TC-NER Pre-Incision Complex / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / mRNA Splicing - Major Pathway / ERKs are inactivated / Initiation of Nuclear Envelope (NE) Reformation / negative regulation of stem cell differentiation / Hydrolases; Acting on ester bonds; Endoribonucleases producing 3'-phosphomonoesters / positive regulation of extrinsic apoptotic signaling pathway in absence of ligand / Beta-catenin phosphorylation cascade / Signaling by GSK3beta mutants / CTNNB1 S33 mutants aren't phosphorylated / CTNNB1 S37 mutants aren't phosphorylated / CTNNB1 S45 mutants aren't phosphorylated / CTNNB1 T41 mutants aren't phosphorylated / RNA polymerase II transcription initiation surveillance / nuclear lumen / Co-stimulation by CD28 / regulation of growth / positive regulation of DNA-templated transcription, elongation / Disassembly of the destruction complex and recruitment of AXIN to the membrane / Abortive elongation of HIV-1 transcript in the absence of Tat / negative regulation of epithelial to mesenchymal transition / inner cell mass cell proliferation / Co-inhibition by CTLA4 / Platelet sensitization by LDL / protein-serine/threonine phosphatase / transcription elongation-coupled chromatin remodeling / RNA Pol II CTD phosphorylation and interaction with CE during HIV infection / RNA Pol II CTD phosphorylation and interaction with CE / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / negative regulation of glycolytic process through fructose-6-phosphate / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / organelle membrane / mRNA Capping / positive regulation of NLRP3 inflammasome complex assembly / ERK/MAPK targets / protein serine/threonine phosphatase activity / mesoderm development / vascular endothelial cell response to oscillatory fluid shear stress / T cell homeostasis / regulation of G1/S transition of mitotic cell cycle / regulation of cell differentiation / regulation of microtubule polymerization / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / phosphoprotein phosphatase activity / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery Similarity search - Function
Journal: Science / Year: 2021 Title: Structural basis of Integrator-mediated transcription regulation. Authors: Isaac Fianu / Ying Chen / Christian Dienemann / Olexandr Dybkov / Andreas Linden / Henning Urlaub / Patrick Cramer / Abstract: Integrator and protein phosphatase 2A (PP2A) form a complex that dephosphorylates paused RNA polymerase II (Pol II), cleaves the nascent RNA, and terminates transcription. We report the structure of ...Integrator and protein phosphatase 2A (PP2A) form a complex that dephosphorylates paused RNA polymerase II (Pol II), cleaves the nascent RNA, and terminates transcription. We report the structure of the pretermination complex containing the human Integrator-PP2A complex bound to paused Pol II. Integrator binds Pol II and the pausing factors DSIF and NELF to exclude binding of the elongation factors SPT6 and PAF1 complex. Integrator also binds the C-terminal domain of Pol II and positions PP2A to counteract Pol II phosphorylation and elongation. The Integrator endonuclease docks to the RNA exit site and opens to cleave nascent RNA about 20 nucleotides from the Pol II active site. Integrator does not bind the DNA clamps formed by Pol II and DSIF, enabling release of DNA and transcription termination.
Mass: 24644.318 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Sus scrofa domesticus (domestic pig) / References: UniProt: I3LSI7
#6: Protein
DNA-directedRNApolymeraseIIsubunitF / DNA-directed RNA polymerases I / II / and III subunit RPABC2 / RPB6 homolog
Mass: 14477.001 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Sus scrofa domesticus (domestic pig) / Tissue: thymus / References: UniProt: F1SKN8
#7: Protein
DNA-directedRNApolymeraseIIsubunitRPB7
Mass: 19314.283 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Sus scrofa domesticus (domestic pig) / References: UniProt: I3LJZ9
#9: Protein
DNA-directedRNApolymeraseIIsubunitRPB9 / RNA polymerase II subunit B9 / DNA-directed RNA polymerase II subunit I / RNA polymerase II 14.5 ...RNA polymerase II subunit B9 / DNA-directed RNA polymerase II subunit I / RNA polymerase II 14.5 kDa subunit / RPB14.5
Mass: 14541.221 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Sus scrofa domesticus (domestic pig) / Tissue: Thymus / References: UniProt: P60899
Mass: 121145.477 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SUPT5H, SPT5, SPT5H / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: O00267
+
Protein/peptide , 2 types, 2 molecules Mu
#13: Protein/peptide
RPBIC-terminaldomainpeptide
Mass: 1356.393 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Sus scrofa domesticus (domestic pig) / Tissue: thymus / References: UniProt: I3LJR4, DNA-directed RNA polymerase
#33: Protein/peptide
Unknown
Mass: 2416.951 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Production host: Trichoplusia ni (cabbage looper)
+
DNA chain , 2 types, 2 molecules NT
#14: DNA chain
Non-templateDNA
Mass: 14712.466 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Human immunodeficiency virus 1
#16: DNA chain
DNATemplate
Mass: 14910.556 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Human immunodeficiency virus 1
+
RNA chain , 1 types, 1 molecules P
#15: RNA chain
TARRNA
Mass: 5410.300 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Human immunodeficiency virus 1
NegativeelongationfactorA / NELF-A / Wolf-Hirschhorn syndrome candidate 2 protein
Mass: 57343.598 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: NELFA, WHSC2, P/OKcl.15 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q9H3P2
#18: Protein
NegativeelongationfactorB / NELF-B / Cofactor of BRCA1
Mass: 68672.789 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: NELFB, COBRA1, KIAA1182 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q8WX92
#19: Protein
NegativeelongationfactorC/D / NELF-C/D / TH1-like protein
Mass: 68544.078 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: NELFCD, NELFD, TH1, TH1L, HSPC130 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q8IXH7
#20: Protein/peptide
NegativeelongationfactorE
Mass: 1890.321 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: NELFE, RD, RDBP / Production host: Trichoplusia ni (cabbage looper)
Mass: 244574.922 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: INTS1, KIAA1440, UNQ1821/PRO3434 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q8N201
#23: Protein
Integratorcomplexsubunit2 / Int2
Mass: 134451.625 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: INTS2, KIAA1287 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q9H0H0
#24: Protein
Integratorcomplexsubunit4 / Int4
Mass: 108306.758 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: INTS4, MSTP093 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q96HW7
#25: Protein
Integratorcomplexsubunit5 / Int5
Mass: 108115.227 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: INTS5, KIAA1698 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q6P9B9
#26: Protein
Integratorcomplexsubunit6 / Int6 / DBI-1 / Protein DDX26 / Protein deleted in cancer 1 / DICE1
Mass: 100527.078 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: INTS6, DBI1, DDX26, DDX26A / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q9UL03
#27: Protein
Integratorcomplexsubunit7 / Int7
Mass: 106952.617 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: INTS7, C1orf73 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q9NVH2
#28: Protein
Integratorcomplexsubunit8 / Int8 / Protein kaonashi-1
Mass: 113219.859 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: INTS8, C8orf52 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q75QN2
#29: Protein
Integratorcomplexsubunit9 / Int9 / Protein related to CPSF subunits of 74 kDa / RC-74
Mass: 73891.219 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: INTS9, RC74 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q9NV88
#30: Protein
Integratorcomplexsubunit11 / Int11 / Cleavage and polyadenylation-specific factor 3-like protein / CPSF3-like protein / Protein ...Int11 / Cleavage and polyadenylation-specific factor 3-like protein / CPSF3-like protein / Protein related to CPSF subunits of 68 kDa / RC-68
Mass: 67770.586 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: INTS11, CPSF3L, RC68 / Production host: Trichoplusia ni (cabbage looper) References: UniProt: Q5TA45, Hydrolases; Acting on ester bonds; Endoribonucleases producing 3'-phosphomonoesters
Mass: 54.938 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Mn / Feature type: SUBJECT OF INVESTIGATION
+
Details
Has ligand of interest
Y
Has protein modification
Y
-
Experimental details
-
Experiment
Experiment
Method: ELECTRON MICROSCOPY
EM experiment
Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction
-
Sample preparation
Component
ID
Name
Type
Entity ID
Parent-ID
Source
1
PEC-Integrator-PP2A complex
COMPLEX
#1-#33
0
MULTIPLESOURCES
2
Pol II-EC
COMPLEX
#1-#16
1
NATURAL
3
human Integrator-PP2A complex
COMPLEX
#22-#33
1
RECOMBINANT
4
NELF
COMPLEX
#17-#20
1
RECOMBINANT
5
DSIF
COMPLEX
#21
1
RECOMBINANT
6
Non-template DNA
COMPLEX
#14
1
MULTIPLESOURCES
7
RNA
COMPLEX
#15
1
RECOMBINANT
8
TemplateDNA
COMPLEX
#16
1
RECOMBINANT
Molecular weight
ID
Entity assembly-ID
Value (°)
Experimental value
1
1
2MDa
NO
2
1
1.5MDa
Source (natural)
ID
Entity assembly-ID
Organism
Ncbi tax-ID
1
2
Sus scrofa domesticus (domestic pig)
9825
2
3
Homo sapiens (human)
9606
3
4
Homo sapiens (human)
9606
4
5
Homo sapiens (human)
9606
5
6
Human immunodeficiency virus 1
32630
6
7
Human immunodeficiency virus 1
32630
7
8
Human immunodeficiency virus 1
32630
Source (recombinant)
ID
Entity assembly-ID
Organism
Ncbi tax-ID
1
3
Trichoplusia ni (cabbage looper)
7111
2
4
Trichoplusia ni (cabbage looper)
7111
3
5
Trichoplusia ni (cabbage looper)
7111
4
6
synthetic construct (others)
32630
5
7
Trichoplusia ni (cabbage looper)
7111
6
8
Trichoplusia ni (cabbage looper)
7111
Buffer solution
pH: 7.4
Specimen
Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
Specimen support
Grid type: Quantifoil R3.5/1
Vitrification
Cryogen name: ETHANE / Humidity: 100 %
-
Electron microscopy imaging
Microscopy
Model: FEI TITAN
Electron gun
Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: OTHER
Electron lens
Mode: OTHER
Image recording
Average exposure time: 2.21 sec. / Electron dose: 46.18 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) / Num. of grids imaged: 1 / Num. of real images: 45000
-
Processing
Software
Name: PHENIX / Classification: refinement
EM software
ID
Name
Version
Category
4
Warp
CTFcorrection
7
UCSF Chimera
1.13
modelfitting
9
PHENIX
1.14
modelrefinement
11
RELION
3.1
finalEulerassignment
12
RELION
classification
13
RELION
3.1
3Dreconstruction
CTF correction
Type: NONE
Particle selection
Num. of particles selected: 4437434
3D reconstruction
Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 614283 / Symmetry type: POINT
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