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Literature summary for 5.6.2.3 extracted from
- G-rich telomeric and ribosomal DNA sequences from the fission yeast genome form stable G-quadruplex DNA structures in vitro and are unwound by the Pfh1 DNA helicase (2016), Nucleic Acids Res., 44, 6213-6231 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of His-FLAG-tagged enzyme in Escherichia coli strain Rosetta 2 (DE3) | Schizosaccharomyces pombe |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K338A | an ATPase-inactive Pfh1 variant | Schizosaccharomyces pombe |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| additional information | presence of G4 structures strongly inhibits the ATPase activity of nPfh1, 5fold inhibition by rDNA G4 oligonucleotide or a single-stranded 57 nt non-G4 oligonucleotide. Phen-DC3-induced G4 structures also inhibit theATP hydrolysis by nPfh1 | Schizosaccharomyces pombe |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| additional information | Pfh1 is enriched at telomeres and rDNAG4 motifs during S phase. rDNA and telomeric G4 structures have different molecularities and are disrupted by mutations in the G-tracts | Schizosaccharomyces pombe | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Schizosaccharomyces pombe |  |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + H2O | Schizosaccharomyces pombe | - |
ADP + phosphate | - |
? | |
| ATP + H2O | Schizosaccharomyces pombe 972 | - |
ADP + phosphate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Schizosaccharomyces pombe | Q9UUA2 | - |
- |
| Schizosaccharomyces pombe 972 | Q9UUA2 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-FLAG-tagged enzyme from Escherichia coli strain Rosetta 2 (DE3) by affinity chromatography | Schizosaccharomyces pombe |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + H2O | - |
Schizosaccharomyces pombe | ADP + phosphate | - |
? | |
| ATP + H2O | - |
Schizosaccharomyces pombe 972 | ADP + phosphate | - |
? | |
| additional information | various oligonucleotides and G4 polymorphic DNA structures are used as substrates. G4 structures are stabilized by the presence of cations, such as Na+ and K+, and the stability and the structure formed can differ depending on the cation species, the ionic strength and the sequence of the oligonucleotide. Determination of G4 consensus motifs from Schizosaccharomyces pombe rDNA and telomeric regions, substrate specificity, detailed overview. nPfh1 unwinds Schizosaccharoyces pombe rDNA and telomeric G4 DNA in vitro, and unwinds Phen-DC3-stabilized G4 structures. The ATP hydrolysis for the telomeric oligonucleotide is lower than for the rDNA oligonucleotide | Schizosaccharomyces pombe | ? | - |
- |
|
| additional information | various oligonucleotides and G4 polymorphic DNA structures are used as substrates. G4 structures are stabilized by the presence of cations, such as Na+ and K+, and the stability and the structure formed can differ depending on the cation species, the ionic strength and the sequence of the oligonucleotide. Determination of G4 consensus motifs from Schizosaccharomyces pombe rDNA and telomeric regions, substrate specificity, detailed overview. nPfh1 unwinds Schizosaccharoyces pombe rDNA and telomeric G4 DNA in vitro, and unwinds Phen-DC3-stabilized G4 structures. The ATP hydrolysis for the telomeric oligonucleotide is lower than for the rDNA oligonucleotide | Schizosaccharomyces pombe 972 | ? | - |
- |
|
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| G4-binding helicase | - |
Schizosaccharomyces pombe |
| Pfh1 | - |
Schizosaccharomyces pombe |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
helicase and ATPase assay at | Schizosaccharomyces pombe |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.5 | - |
helicase and ATPase assay at | Schizosaccharomyces pombe |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | hPIF1 belongs to the evolutionarily conserved Pif1 family, which comprises multifunctional helicases with a unique 21 amino acid signature motif. Pfh1 is the sole Sschizosaccharomyces pombe Pif1 member | Schizosaccharomyces pombe |
| malfunction | presence of G4 structures strongly inhibits the ATPase activity of nPfh1 | Schizosaccharomyces pombe |
| metabolism | several human helicases, such as WRN, FANCJ, BLM and PIF1 (hPIF1), are G4-binding helicases that are linked to genetic diseases. For example, the WRN helicase is associated with Werner syndrome, FANCJ with Fanconi anemia and hPIF1 with familial breast cancer | Schizosaccharomyces pombe |
| physiological function | G-rich telomeric and ribosomal DNA sequences from the fission yeast genome form stable G-quadruplex DNA structures in vitro and are unwound by the Pfh1 DNA helicase, as well as G4 motifs from Schizosaccharoyces pombe rDNA and , that form predominantly inter- and intramolecular G4 structures, respectively. Mutations in the G-tracts of rDNA and telomeric G4 DNA oligonucleotides inhibit the formation of G4 structures | Schizosaccharomyces pombe |
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