Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | Brr2 can be autoinhibited via a large N-terminal region folding back onto its helicase core and autoactivated by a catalytically inactive C-terminal helicase cassette | Homo sapiens | |
additional information | Brr2 can be autoinhibited via a large N-terminal region folding back onto its helicase core and autoactivated by a catalytically inactive C-terminal helicase cassette | Thermochaetoides thermophila | |
Prp8 protein | the Jab1 domain of the Prp8 protein can inhibit Brr2 by intermittently inserting a C-terminal tail in the enzyme's RNA-binding tunnel or activate the helicase after removal of this tail. Binding of the Prp8 Jab1 C-terminal tail at the Brr2 RNA binding tunnel is evolutionarily conserved, Brr2-Jab1 binding structure and analysis, overview | Homo sapiens | |
Prp8 protein | the Jab1 domain of the Prp8 proten can inhibit Brr2 by intermittently inserting a C-terminal tail in the enzyme's RNA-binding tunnel or activate the helicase after removal of this tail. Binding of the Prp8 Jab1 C-terminal tail at the Brr2 RNAbinding tunnel is evolutionarily conserved, Brr2-Jab1 binding structure and analysis, overview | Thermochaetoides thermophila |
Cloned (Comment) | Organism |
---|---|
gene BRR2, recombinant expression of His10-tagged TEV-cleavable codon-optimized DNA fragments encoding selected regions of yBrr2 in Escherichia coli strain Rosetta2 (DE3) | Homo sapiens |
gene CTHT_0009470, recombinant expression of His10-tagged TEV-cleavable codon-optimized DNA fragments encoding selected regions of cBrr2 in Escherichia coli strain Rosetta2 (DE3) | Thermochaetoides thermophila |
gene SNRNP200, recombinant expression of His10-tagged TEV-cleavable codon-optimized DNA fragments encoding selected regions of hBrr2 in Escherichia coli strain Rosetta2 (DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant Brr2-Jab1 complex, for complex 1: yBrr2FL and yJab1 are mixed in a 1:2 molar ratio in 10 mM Tris-HCl, pH 7.5, 150 mM NaCl, 2 mM DTT, and separated by gel filtration and and concentrated to 2 mg/ml, followed by sitting drop vapor diffusion technique, mixing of 0.0013 ml of protein solution with 0.0013 ml of reservoir solution containing 0.1 M Tris-HCl, pH 7.5, 10.5% w/v PEG 3350, and 0.2 M MgCl2, 18°C. For complex 2: yBrr2T2, yJab1 and yNtr2 are mixed in a 1:5:5 molar ratio in 10 mM Tris-HCl, pH 7.5, 150 mM NaCl, 2 mM DTT, and separated by gel filtration and and concentrated to 4 mg/ml, followed by hanging drop vapor diffusion technique, mixing of 0.0005 ml of protein solution with 0.00025 ml of reservoit solution containing 0.1 M MES-NaOH, pH 6.5, 9.2% w/v PEG 4000, 0.4 M MgCl2, and with 0.00025 ml 0.33% w/v 1,5-naphthalenedisulfonic acid, 18°C. X-ray diffraction structure determination and analysis at 3.4-4.2 A resolution, molecular replacement using the yBrr2T4-Jab1 structure coordinates as the search model (PDB ID 4BGD) | Homo sapiens |
purified recombinant Brr2-Jab1 complex, X-ray diffraction structure determination and analysis at 3.2 A resolution | Thermochaetoides thermophila |
purified recombinant Brr2-Jab1 complex, X-ray diffraction structure determination and analysis at 3.4-4.2 A resolution | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
additional information | generation of codon-optimized DNA fragments encoding selected regions of hBrr2 | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | Brr2 can be autoinhibited via a large N-terminal region folding back onto its helicase core and autoactivated by a catalytically inactive C-terminal helicase cassette | Homo sapiens | |
additional information | Brr2 can be autoinhibited via a large N-terminal region folding back onto its helicase core and autoactivated by a catalytically inactive C-terminal helicase cassette. Effects of NTR truncations on Chaetomium thermophilum Brr2NC variants. Binding of N-terminal Brr2NC truncations to U4/U6 di-snRNA monitored by electrophoretic mobility shift assays and time courses of U4/U6 unwinding by Brr2NC variants, overview | Thermochaetoides thermophila | |
Prp8 protein | the Jab1 domain of the Prp8 proten can inhibit Brr2 by intermittently inserting a C-terminal tail in the enzyme's RNA-binding tunnel or activate the helicase after removal of this tail. Binding of the Prp8 Jab1 C-terminal tail at the Brr2 RNA binding tunnel is evolutionarily conserved, Brr2-Jab1 binding structure and analysis, overview. Jab1-based inhibition of Brr2 presumably takes effect in all eukaryotes but is implemented via organism-specific molecular contacts. Brr2 autoinhibition can act in concert with Jab1-mediated inhibition. The NTR and the Jab1 tail cooperate in inhibiting RNA binding by Brr2; the Jab1 domain of the Prp8 proten can inhibit Brr2 by intermittently inserting a C-terminal tail in the enzyme's RNA-binding tunnel or activate the helicase after removal of this tail. Binding of the Prp8 Jab1 C-terminal tail at the Brr2 RNA binding tunnel is evolutionarily conserved, Brr2-Jab1 binding structure and analysis, overview. Jab1-based inhibition of Brr2 presumably takes effect in all eukaryotes but is implemented via organism-specific molecular contacts. Brr2 autoinhibition can act in concert with Jab1-mediated inhibition.The NTR and the Jab1 tail cooperate in inhibiting RNA binding by Brr2 | Homo sapiens | |
Prp8 protein | the Jab1 domain of the Prp8 proten can inhibit Brr2 by intermittently inserting a C-terminal tail in the enzyme's RNA-binding tunnel or activate the helicase after removal of this tail. Binding of the Prp8 Jab1 C-terminal tail at the Brr2 RNA binding tunnel is evolutionarily conserved, Brr2-Jab1 binding structure and analysis, overview. Jab1-based inhibition of Brr2 presumably takes effect in all eukaryotes but is implemented via organism-specific molecular contacts. Brr2 autoinhibition can act in concert with Jab1-mediated inhibition.The NTR and the Jab1 tail cooperate in inhibiting RNA binding by Brr2 | Thermochaetoides thermophila |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Homo sapiens | |
Mg2+ | required | Thermochaetoides thermophila |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | Homo sapiens | - |
ADP + phosphate | - |
? | |
ATP + H2O | Thermochaetoides thermophila | - |
ADP + phosphate | - |
? | |
ATP + H2O | Thermochaetoides thermophila IMI 039719 | - |
ADP + phosphate | - |
? | |
ATP + H2O | Thermochaetoides thermophila DSM 1495 | - |
ADP + phosphate | - |
? | |
ATP + H2O | Thermochaetoides thermophila CBS 144.50 | - |
ADP + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | O75643 | - |
- |
Thermochaetoides thermophila | G0S0B9 | - |
- |
Thermochaetoides thermophila CBS 144.50 | G0S0B9 | - |
- |
Thermochaetoides thermophila DSM 1495 | G0S0B9 | - |
- |
Thermochaetoides thermophila IMI 039719 | G0S0B9 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His10-tagged Brr2 fragments from Escherichia coli strain Rosetta2 (DE3) by nickel affinity chromatography, tag cleavage by TEV protease, and gel filtration | Homo sapiens |
recombinant His10-tagged Brr2 fragments from Escherichia coli strain Rosetta2 (DE3) by nickel affinity chromatography, tag cleavage by TEV protease, and gel filtration | Thermochaetoides thermophila |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | - |
Homo sapiens | ADP + phosphate | - |
? | |
ATP + H2O | - |
Thermochaetoides thermophila | ADP + phosphate | - |
? | |
ATP + H2O | - |
Thermochaetoides thermophila IMI 039719 | ADP + phosphate | - |
? | |
ATP + H2O | - |
Thermochaetoides thermophila DSM 1495 | ADP + phosphate | - |
? | |
ATP + H2O | - |
Thermochaetoides thermophila CBS 144.50 | ADP + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BRR2 | - |
Homo sapiens |
BRR2 | - |
Thermochaetoides thermophila |
CTHT_0009470 | - |
Thermochaetoides thermophila |
SNRNP200 | - |
Homo sapiens |
spliceosomal RNA helicase | - |
Homo sapiens |
spliceosomal RNA helicase | - |
Thermochaetoides thermophila |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
helicase assay at | Homo sapiens |
30 | - |
helicase assay at | Thermochaetoides thermophila |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
helicase assay at | Homo sapiens |
7.5 | - |
helicase assay at | Thermochaetoides thermophila |
General Information | Comment | Organism |
---|---|---|
evolution | binding of the Prp8 Jab1 C-terminal tail at the Brr2 RNA binding tunnel is evolutionarily conserved | Homo sapiens |
evolution | binding of the Prp8 Jab1 C-terminal tail at the Brr2 RNA binding tunnel is evolutionarily conserved | Thermochaetoides thermophila |
physiological function | RNA helicase Brr2 is implicated in multiple phases of pre-mRNA splicing and thus requires tight regulation. Interplay of cis- and trans-regulatory mechanisms in the spliceosomal RNA helicase Brr2. Brr2 can be autoinhibited via a large N-terminal region folding back onto its helicase core and autoactivated by a catalytically inactive C-terminal helicase cassette. It can be regulated in trans by the Jab1 domain of the Prp8 protein, which can inhibit Brr2 by intermittently inserting a C-terminal tail in the enzyme's RNA-binding tunnel or activate the helicase after removal of this tail. Brr2 autoinhibition can act in concert with Jab1-mediated inhibition. The N-terminal region influences how the Jab1 C-terminal tail interacts at the RNA-binding tunnel. the N-terminal region and the Jab1 C-terminal tail specifically interfere with accommodation of double-stranded and single-stranded regions of an RNA substrate, respectively, mutually reinforcing each other. Regulation based on the N-terminal region requires the presence of the inactive C-terminal helicase cassette, intricate system of regulatory mechanisms, which control Brr2 activities during snRNP assembly and splicing, overview | Homo sapiens |
physiological function | RNA helicase Brr2 is implicated in multiple phases of pre-mRNA splicing and thus requires tight regulation. Interplay of cis- and trans-regulatory mechanisms in the spliceosomal RNA helicase Brr2. Brr2 can be autoinhibited via a large N-terminal region folding back onto its helicase core and autoactivated by a catalytically inactive C-terminal helicase cassette. It can be regulated in trans by the Jab1 domain of the Prp8 protein, which can inhibit Brr2 by intermittently inserting a C-terminal tail in the enzyme's RNA-binding tunnel or activate the helicase after removal of this tail. Brr2 autoinhibition can act in concert with Jab1-mediated inhibition. The N-terminal region influences how the Jab1 C-terminal tail interacts at the RNA-binding tunnel. the N-terminal region and the Jab1 C-terminal tail specifically interfere with accommodation of double-stranded and single-stranded regions of an RNA substrate, respectively, mutually reinforcing each other. Regulation based on the N-terminal region requires the presence of the inactive C-terminal helicase cassette, intricate system of regulatory mechanisms, which control Brr2 activities during snRNP assembly and splicing, overview | Thermochaetoides thermophila |