Recombinant Saccharomyces cerevisiae DNA topoisomerase 1 (TOP1), partial

Code CSB-YP024056SVG
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Source Yeast
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Code CSB-EP024056SVG
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Source E.coli
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Code CSB-EP024056SVG-B
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Source E.coli
Conjugate Avi-tag Biotinylated
E. coli biotin ligase (BirA) is highly specific in covalently attaching biotin to the 15 amino acid AviTag peptide. This recombinant protein was biotinylated in vivo by AviTag-BirA technology, which method is BriA catalyzes amide linkage between the biotin and the specific lysine of the AviTag.
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Code CSB-BP024056SVG
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Source Baculovirus
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Code CSB-MP024056SVG
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Source Mammalian cell
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Product Details

Purity
>85% (SDS-PAGE)
Target Names
TOP1
Uniprot No.
Alternative Names
TOP1; MAK1; YOL006C; DNA topoisomerase 1; EC 5.6.2.1; DNA topoisomerase I; Maintenance of killer protein 1
Species
Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast)
Protein Length
Partial
Tag Info
Tag type will be determined during the manufacturing process.
The tag type will be determined during production process. If you have specified tag type, please tell us and we will develop the specified tag preferentially.
Form
Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer before Lyophilization
Tris/PBS-based buffer, 6% Trehalose, pH 8.0
Reconstitution
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20℃/-80℃. Our default final concentration of glycerol is 50%. Customers could use it as reference.
Troubleshooting and FAQs
Storage Condition
Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
Shelf Life
The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Lead Time
Delivery time may differ from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Note: All of our proteins are default shipped with normal blue ice packs, if you request to ship with dry ice, please communicate with us in advance and extra fees will be charged.
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Datasheet
Please contact us to get it.

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Target Background

Function
Releases the supercoiling and torsional tension of DNA introduced during the DNA replication and transcription by transiently cleaving and rejoining one strand of the DNA duplex. Introduces a single-strand break via transesterification at a target site in duplex DNA. The scissile phosphodiester is attacked by the catalytic tyrosine of the enzyme, resulting in the formation of a DNA-(3'-phosphotyrosyl)-enzyme intermediate and the expulsion of a 5'-OH DNA strand. The free DNA strand then rotates around the intact phosphodiester bond on the opposing strand, thus removing DNA supercoils. Finally, in the religation step, the DNA 5'-OH attacks the covalent intermediate to expel the active-site tyrosine and restore the DNA phosphodiester backbone.
Gene References into Functions
  1. Studies demonstrate opposing roles of Top1 activity in either promoting or destabilizing genome integrity. Resolution of transcription-associated negative supercoils, for example, prevents pairing of the nascent RNA with the DNA template (R-loops) as well as DNA secondary structure formation. Reduced Top1 levels thus enhance CAG repeat contraction, somatic hypermutation, and class switch recombination. PMID: 29177731
  2. Our findings reveal a functional interplay between Mms21 and Top1 in maintenance of longer chromosomes, and suggest that lack of sumoylation of Mms21 targets coupled with Top1 deficiency is a crucial requirement for accurate inheritance of longer chromosomes. PMID: 27872982
  3. Too much, as well as too little of Top1 is detrimental to eukaryotic genomes, and that CPT has destabilizing effects that extend beyond those associated with DSB formation. PMID: 28961461
  4. The authors demonstrate biochemically that irreversible double-strand breaks are generated by subsequent Top1 cleavage on the opposite strand from the Top1-induced DNA nicks at ribonucleotide sites. PMID: 27932446
  5. The role of topoisomerase I in suppressing genome instability associated with a highly transcribed guanine-rich sequence is not restricted to preventing RNA:DNA hybrid accumulation. PMID: 26527723
  6. top1 mutants had greatly elevated levels of instability within the tandemly-repeated ribosomal RNA genes. PMID: 25830313
  7. Top1 is a critical factor in suppression of instability associated with potential G4 DNA-forming sequences. PMID: 25473964
  8. analysis of how human and yeast DNA topoisomerase I domain interactions impact enzyme activity and sensitivity to camptothecin PMID: 25795777
  9. Top1 reversibly cleaves genomic ribonucleotides with formation of a ribonucleotide 2',3'-cyclic phosphate.Top1 cleaves two nucleotides upstream of the ribonucleotide 2',3'-cyclic phosphate to yield a covalent Top1-DNA complex. PMID: 25777529
  10. Top1-dependent deletions are initiated either by Top1 cleavage complex trapping or by Top1 incision at ribonucleoside monophosphates in DNA. PMID: 23305949
  11. study demonstrates that rNMP (ribonucleoside monophosphates)-associated deletion formation requires the activity of Top1; studies extend the role of Top1 to include the processing of rNMPs in genomic DNA into irreversible single-strand breaks PMID: 21700875
  12. analysis of unavoidable conflicts between Top1p and the transcriptional machinery and their potential impact on genome stability PMID: 21177431
  13. Loss of Top1 enhances inherent R-loop formation, particularly over the 5' region of the rDNA, imposing persistent transcription blocks when RNase H is limiting. PMID: 20634320
  14. independent alterations in DNA cleavage and religation contribute to the lethal phenotype of DNA topoisomerase I active site mutations PMID: 15489506
  15. Data suggest that topoisomerase II is the main modulator of DNA topology in chromatin fibers, and that topoisomerase I then assists DNA relaxation where chromatin structure impairs DNA juxtaposition but allows twist diffusion. PMID: 16710299
  16. Changes in the flexibility or orientation of the linker alter the geometry of the active site and thereby the kinetics of DNA cleavage/religation. PMID: 17276985
  17. data are consistent with a coordinated action of Top1 and Top2 in counteracting the accumulation of torsional stress and sister chromatid entanglement at replication forks PMID: 17671091
  18. Top1p can optimize transcript levels for cell growth in exponentially growing cells under a synthetic medium with glucose PMID: 18272174
  19. Disulfide cross-links reveal conserved features of DNA topoisomerase I architecture and a role for the N terminus in clamp closure PMID: 18693244

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Subcellular Location
Nucleus, nucleolus. Nucleus, nucleoplasm.
Protein Families
Type IB topoisomerase family
Database Links

KEGG: sce:YOL006C

STRING: 4932.YOL006C

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