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Literature summary for 3.6.4.13 extracted from

  • Absmeier, E.; Becke, C.; Wollenhaupt, J.; Santos, K.F.; Wahl, M.C.
    Interplay of cis- and trans-regulatory mechanisms in the spliceosomal RNA helicase Brr2 (2017), Cell Cycle, 16, 100-112 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.6.4.13

Activating Compound

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(Commentary)

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 (Commentary)

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

Protein Variants Comment Organism
additional information generation of codon-optimized DNA fragments encoding selected regions of hBrr2 Homo sapiens

Inhibitors

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

Metals/Ions Comment Organism Structure
Mg2+ required Homo sapiens
Mg2+ required Thermochaetoides thermophila

Natural Substrates/ Products (Substrates)

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

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 (Commentary)

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 and Products (Substrate)

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 helicase assay at Homo sapiens
30
-
 helicase assay at Thermochaetoides thermophila

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 helicase assay at Homo sapiens
7.5
-
 helicase assay at Thermochaetoides thermophila

General Information

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