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- PDB-2yh1: Model of human U2AF65 tandem RRM1 and RRM2 domains with eight-sit... -
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Open data
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Basic information
Entry | Database: PDB / ID: 2yh1 | ||||||
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Title | Model of human U2AF65 tandem RRM1 and RRM2 domains with eight-site uridine binding | ||||||
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![]() | TRANSCRIPTION / PRE-MRNA SPLICING | ||||||
Function / homology | ![]() U2AF complex / poly-pyrimidine tract binding / pre-mRNA 3'-splice site binding / mRNA 3'-end processing / C2H2 zinc finger domain binding / commitment complex / Transport of Mature mRNA derived from an Intron-Containing Transcript / RNA Polymerase II Transcription Termination / U2-type prespliceosome / molecular function inhibitor activity ...U2AF complex / poly-pyrimidine tract binding / pre-mRNA 3'-splice site binding / mRNA 3'-end processing / C2H2 zinc finger domain binding / commitment complex / Transport of Mature mRNA derived from an Intron-Containing Transcript / RNA Polymerase II Transcription Termination / U2-type prespliceosome / molecular function inhibitor activity / spliceosomal complex assembly / Protein hydroxylation / negative regulation of mRNA splicing, via spliceosome / negative regulation of protein ubiquitination / mRNA Splicing - Major Pathway / positive regulation of RNA splicing / spliceosomal complex / mRNA splicing, via spliceosome / mRNA processing / nuclear speck / enzyme binding / RNA binding / nucleoplasm / nucleus Similarity search - Function | ||||||
Biological species | ![]() | ||||||
Method | SOLUTION NMR / MODIFIED ARIA | ||||||
![]() | Mackereth, C.D. / Madl, T. / Simon, B. / Zanier, K. / Gasch, A. / Sattler, M. | ||||||
![]() | ![]() Title: Multi-Domain Conformational Selection Underlies Pre-Mrna Splicing Regulation by U2Af Authors: Mackereth, C.D. / Madl, T. / Bonnal, S. / Simon, B. / Zanier, K. / Gasch, A. / Rybin, V. / Valcarcel, J. / Sattler, M. #1: ![]() Title: Structural Basis for Polypyrimidine Tract Recognition by the Essential Pre-Mrna Splicing Factor U2Af65. Authors: Sickmier, E.A. / Frato, K.E. / Shen, H. / Paranawithana, S.R. / Green, M.R. / Kielkopf, C.L. | ||||||
History |
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Structure visualization
Structure viewer | Molecule: ![]() ![]() |
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Downloads & links
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PDBx/mmCIF format | ![]() | 701.8 KB | Display | ![]() |
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PDB format | ![]() | 597.9 KB | Display | ![]() |
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-Validation report
Arichive directory | ![]() ![]() | HTTPS FTP |
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-Related structure data
Related structure data | ![]() 2yh0C ![]() 2yh4 C: citing same article ( |
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Similar structure data | |
Other databases |
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Links
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Assembly
Deposited unit | ![]()
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NMR ensembles |
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Components
#1: Protein | Mass: 21510.525 Da / Num. of mol.: 1 Fragment: TANDEM RRM1 AND RRM2 DOMAINS JOINED BY NATIVE LINKER, RESIDUES 148-342 Source method: isolated from a genetically manipulated source Details: STRUCTURE BASED ON DATA FROM TEN CYSTEINE MUTANTS (N155C, A164C, A171C, L187C, A188C, T209C, D273C, S281C, A287C AND A318C) COVALENTLY MODIFIED BY 3-(2-IODOACETAMIDO)-2,2,5,5, TETRAMETHYL-1- ...Details: STRUCTURE BASED ON DATA FROM TEN CYSTEINE MUTANTS (N155C, A164C, A171C, L187C, A188C, T209C, D273C, S281C, A287C AND A318C) COVALENTLY MODIFIED BY 3-(2-IODOACETAMIDO)-2,2,5,5, TETRAMETHYL-1-PYRROLIDINYLOXY RADICAL (IODOACETAMIDO-PROXYL) Source: (gene. exp.) ![]() ![]() ![]() |
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#2: RNA chain | Mass: 2710.535 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) ![]() |
-Experimental details
-Experiment
Experiment | Method: SOLUTION NMR | ||||||||||||||||||||||||
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NMR experiment |
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NMR details | Text: THE STRUCTURE WAS DETERMINED USING A MULTI-STEP PROTOCOL AS DESCRIBED IN REMARK 3. |
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Sample preparation
Details | Contents: 90% WATER/10% D2O | ||||||||||||||||||||
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Sample conditions |
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-NMR measurement
NMR spectrometer |
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Processing
NMR software |
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Refinement | Method: MODIFIED ARIA / Software ordinal: 1 Details: STRUCTURE CALCULATION INCLUDED DISTANCE RESTRAINTS BASED ON PARAMAGNETIC RELAXATION ENHANCEMENT DATA FROM TEN INDEPENDENT CYSTEINE MUTANTS (N155C, A164C, A171C, L187C, A188C, T209C, D273C, ...Details: STRUCTURE CALCULATION INCLUDED DISTANCE RESTRAINTS BASED ON PARAMAGNETIC RELAXATION ENHANCEMENT DATA FROM TEN INDEPENDENT CYSTEINE MUTANTS (N155C, A164C, A171C, L187C, A188C, T209C, D273C, S281C, A287C AND A318C) COVALENTLY MODIFIED BY 3-(2-IODOACETAMIDO)-2,2,5,5, TETRAMETHYL-1-PYRROLIDINYLOXY RADICAL (IODOACETAMIDO- PROXYL). THE STRUCTURE WAS DETERMINED USING A MULTI-STEP PROTOCOL. FIRST WITH REFINEMENT OF INDIVIDUAL STRUCTURED DOMAINS FOR RRM1 AND RRM2 OF U2AF65. THE INITIAL COORDINATES OF THE ISOLATED DOMAINS WERE BASED ON PDB ENTRY 2G4B BY SICKMIER ET AL. 2006 (SEE REMARK 1 REFERENCE). SECOND STEP WAS ADDITION AND RANDOMIZATION OF FLEXIBLE AND LINKER RESIDUES. THE THIRD STEP WAS MODEL CALCULATION USING RDC ORIENTATION, PRE-BASED DISTANCE, TALOS DIHEDRAL AND HYDROGEN BOND RESTRAINTS WITH THE RRM1 AND RRM2 DOMAINS RESTRAINED TO THEIR INITIAL STARTING STRUCTURES. INTER- MOLECULAR PROTEIN-RNA DISTANCE RESTRAINTS WERE BASED ON ATOMIC DETAILS FROM PDB ENTRY 2G4B AND ASSUMED AN EIGHT- URIDINE BINDING MODE FOR U2AF65 RRM1-RRM2. | ||||||||||||
NMR ensemble | Conformer selection criteria: LOWEST ENERGY / Conformers calculated total number: 125 / Conformers submitted total number: 10 |