The phosphorylation of Drosophila melanogaster DNA topoisomerase II by purified casein kinase II was characterized in vitro. Topoisomerase II InhibitorsAzacyanines as Novel Topoisomerase II Alpha Inhibitors Such a type II enzyme has also been characterized in Trypanosoma equiperdum. What is Topoisomerase II? (2002). They are broadly classified into two categories namely, Type II A and Type IIB. Topo II activity is a suggested predictive marker in cancer therapy and, consequently, the described highly sensitive monitoring of Topo II activity may add considerably to the toolbox of individualized medicine where decisions are based on very sparse samples. Jinlin Li, Noemi Nagy, Jiangnan Liu, Soham Gupta, Teresa Frisan, Thomas Hennig, Donald P. Cameron, Laura Baranello, Maria G. Masucci The purified topoisomerase had a specific activity of 5 x 105 units/mg, in which1 unitofactivityis definedastheamount of enzyme required to unknot 0.3 ,g of P4 knotted DNA underthe standard assay conditions. 2021; 10 : e65184 Crossref Background: The substituted 1,8-Naphthalimides (1H-benzo [de]isoquinoline-1,3 (2H)- diones) are known as DNA intercalators stabilizing DNA-Topoisomerase II complexes. DNA sequence analysis revealed an ORF of 3711 bp encoding a with protein led us to investigate the involvement of topoisomerase II (topo II) in their formation. Type II topoisomerase change the linking number of circular DNA by ± 2. There are several reports in the literature that implicate reduced topoisomerase II activity in the drug resistance phenotype (19, 31-37). 0.5 mM ATP, 0.5 mM dithiothreitol) with 0.2 µg KDNA. [Source 12)] Topoisomerase 2. II. Doxorubicin (Dox) is the first-line drug for the TNBC treatment, acting as a DNA intercalator and topoisomerase II (Topo II) inhibitor; however, it has been observed to exhibit strong cardiotoxicity. (ii) ATP independent relaxation activity was not detected. Topoisomerase poisons, in contrast to topoisomerase catalytic inhibitors, often have good anticancer activity. NA topoisomerase II (TopoII ) is an essential chromosome-associated enzyme with activity im-plicated in the resolution of tangled DNA at cen-tromeres before anaphase onset. Here, we show that PIASy-mediated small ubiquitin-like modifier Schmidt, B., Osheroff, N. & Berger, J. Download Download PDF. Decatenation assays are carried out using kinetoplast DNA substrate in a final volume of 20-30 µl in topo II reaction buffer (50 mM Tris-Cl, pH 8.0, 120 mM KCl, 10 mM MgCl2. Topoisomerase II inhibitors 1 July 2011 Topoisomerase II Inhibitors Topoisomerase II inhibitors are chemicals that inhibit a group of DNA enzymes called type II topoisomerases (topoisomerase IIs). Read Paper. Ginkgo biloba leaf extract induces DNA damage by inhibiting topoisomerase II activity in human hepatic cells. Furthermore, we see a 100-fold decrease in the LD50 of etoposide, another topo II inhibitor. Raji-HN2 cells contained -3 times as much enzyme activity Topoisomerase I and II activity co-segregated in individual colon tumour samples, but no correlation was observed in cervix, lung or breast tumours. Rather, the activity of the topoisomerase II inhibitors was unexpectedly antagonized. At last one final time, the enzyme opens up and releases the active site and move to another site. Initial work indicates that the enzyme interacts with specific forms of DNA damage and that lesions alter topoisomerase II activity in a manner similar to that of anticancer drugs. Topoisomerase II (TOP2) has held the interest of researchers studying cancer owing to the discovery that it is targeted by active anticancer drugs, notably etoposide and doxorubicin (Ref. eLife. In eukaryotes, IIA enzymes catalyze the relaxation of positively or negatively supercoiled DNA, as well as the decatenation and unknotting of DNA helices. Purpose: Topoisomerase IIα content, topoisomerase II catalytic activity and drug sensitivities to the topoisomerase II inhibitors, doxorubicin and etoposide, were examined in a panel of 14 unselected human lung cancer cell lines in order to determine the relationship between topoisomerase II and drug sensitivities to the topoisomerase II inhibitors. DNA Topoisomerase II Structure. Phosphorylation stimulated the DNA relaxation activity of topoisomerase II by 3-fold over that of the dephosphorylated enzyme, and the effects of modification could be reversed by treatment with alkaline phosphatase. The drug has activity as monotherapy in recurrent acute myelogenous leukemia (AML) and in platinum-refractory ovarian cancer. Laura Baranello. link formation. RNA polymerase II regulates topoisomerase 1 activity to favor efficient transcription. The tyrosine residue of activated topoisomerase attacks directly the phosphodiester bonds of broken DNA. The tyrosine is now bound with the phosphate of the broken DNA strand at -5′ end. The other -3’OH end remains free and held by the topoisomerase domain. Recent studies in budding yeast have shown that some mechanism minimizes the knotting probability of intracellular DNA. Cell 165 , 357–371 (2016). Material and Methods They are broadly classified into two Topoisomerase-II activity was analyzed in various human leukemic and lymphoblastoid cell-lines with comparison to normal human peripheral blood lymphocytes. 2-propionyl-thiazole backbone, and their Cu(II) complexes, and shows that the Cu complexes not only inhibit function of human topoisomerase IIα, but have significant activity against two different breast cancer cell lines. … Addition of purified topoisomerase II-alpha reconstituted chromatin-dependent activation activity in reactions with core pol II. by targeting proteins that modulate topoisomerase II activity. 0 1996 Cold Spring Harbor Laboratory Press 0-87969-459-9/96 $5 + .OO 587. Baranello, L. et al. They need ATP hydrolyzing for this activity. The role of DNA topoisomerases (Topo) IIα and IIβ was investigated in various drug-resistant melanoma cells. Human recombinant TOP1 and BRD4 were purified and Levens, D. (2014). A human Burkitt lymphoma cell line, Raji-HN2, made 10-fold more resistant to nitrogen mustard (HN2) than the parental Raji cell line, exhibited the following characteristics when compared to the parental Raji cells: (i) decreased HN2-induced DNA interstrand crosslinking; (ii) increased (3-fold) DNA topoisomerase II [DNA topoisomerase (ATP-hydrolyzing), EC 5.99.1.3] activity; (iii) … Type II DNA topoisomerases are tetrameric proteins formed by two different subunits, GyrA2GyrB2 for gyrase and ParC2ParE2 for DNA topoisomerase IV 13).Requiring ATP, these enzymes act by making a transient break on the double stranded DNA, passing through an intact duplex DNA via the broken strand followed by a resealing of the … It has been shown that Topoisomerase II activity is required for HIV-1 replication and the enzyme is phosphorylated during early time points of HIV-1 replication. Type II topoisomerase change the linking number of circular DNA by ± 2. TAS-103 is a recently developed dual inhibitor of topoisomerase-I (topo-I) and topoisomerase-II (topo-II). View. In order to manage the DNA tangles and supercoils, the type II topoisomerase cut both the DNA strands simultaneously. Topoisomerase II Activity. A short summary of this paper. Topoisomerase II, extracted from nuclei with 0.35 M NaCI, was assayed by the P4 DNA unknotting assay (Fig. In recent years, a great interest has emerged in resveratrol (RSV) activity in the prevention of various pathologies including cancer. The main difference between Topoisomerase I and II is that topoisomerase I cut one strand of the DNA double helix whereas topoisomerase II cut both strands of the DNA double helix.Furthermore, topoisomerase I do not require ATP hydrolysis while topoisomerase II requires ATP hydrolysis. This effect is likely related to the multiplicity of genes regulated by the Stat3 transcription factor, including the cell cycle regulatory protein, cyclin D1, in addition to genes like Bcl-x L that control apoptosis. One of these, heat shock protein-90 (Hsp90), is itself a Name Topoisomerase II Inhibitors Accession Number DBCAT000549 Description. Voreloxin is a naphthyridine derivative that acts partly by inhibition of topoisomerase II, causing replication-dependent DNA damage in the S-phase of the cell cycle and leading to apoptosis via irreversible G2 arrest. Topoisomerase IIs regulate the structure of DNA and are essential in separating multiple intertwined DNA daughter strands after DNA replication and prior to mitosis. These enzymes resolve knots and tangles in the genetic material by transiently creating and resealing DNA double strand breaks. The intact holoenzyme is a 97 kDa protein. The Ecoli topoisomerase I is a holoenzyme with three Zn (II) atoms in the tetracysteine motifs near its carboxy terminus. It is 97 kDa in weight. Topoisomerase I has several unusual features. It does not need ATP hydrolyzing to catalyze the topological rearrangement of DNA. CAS PubMed PubMed Central Google Scholar The most potent derivative, 32, was shown to induce the apoptosis pathway. Under the conditions used, the kinase incorporated a maximum of 2-3 molecules of phosphate per homodimer of topoisomerase II. In humans, … In the present study, andrographolide and its structurally modified analogues were investigated for their inhibitory activities on hTopo II α enzyme. Inhibition of Topoisomerase II DNA Relaxation Activity. Topoisomerase is an enzyme tasked with cutting, or influencing DNA to repair breakage and avoid further complication when necessary. It has also been shown to alkylate DNA through an epoxide-mediated electrophilic attack, and this alkylation is greatly enhanced at specific sites by topoisomerase II. The obtained results highlight 3-methyl-2-phenyl-1H-indole as a The juxtaposition of intracellular DNA segments, together with the DNA-passage activity of topoisomerase II, leads to the formation of DNA knots and interlinks, which jeopardize chromatin structure and gene expression. Topoisomerase II (topo II) is essential for disentangling newly replicated chromosomes. The main difference between Topoisomerase I and II is that topoisomerase I cut one strand of the DNA double helix whereas topoisomerase II cut both strands of the DNA double helix.Furthermore, topoisomerase I do not require ATP hydrolysis while topoisomerase II requires ATP hydrolysis. Int. Because topoisomerase II is postulated to play a role in DNA repair (14), it was of interest to compare the topoisomerase II activity of Raji and Raji-HN2 cells. Topoisomerases (or DNA topoisomerases) are enzymes that participate in the overwinding or underwinding of DNA. The enzymes involved in DNA metabolism may offer targets for chemotherapy, especially in rapidly proliferating cells. Like topoisomerase I, topoisomerase II catalyzes relaxation of supercoiled DNA. The structural representations for these new compounds are shown in Figure 1. TAS-103 has documented cytotoxicity in vitro and antitumor activity against a variety of mouse, rat, and human xenografts in vivo. Topoisomerase I has also been implicated in knotting and unknotting DNA (1) and in linking complementary rings of single-stranded DNA into double-stranded rings (2). Samples were assembled in ice in 50 mmol/L Tris-HCl (pH 8.0), 120 mmol/L KCl, 10 mmol/L MgCl 2, 0.5 mmol/L DTT, 30 μg/mL bovine serum albumin, and 0.5 mmol/L ATP prepared extemporarily. et al. Thus, a topological change on DNA drives topoisomerase II to decatenate molecules during mitosis, potentially driving the full decatenation of the genome. The monocyclic 1,4-benzoquinone, HU-331, the direct oxidation product of cannabidiol, inhibits the catalytic activity of topoisomerase II but without inducing DNA strand breaks or generating free radicals, and unlike many … Topoisomerase IIs regulate the structure of DNA and are essential in separating multiple intertwined DNA daughter strands after The winding problem of DNA arises due to the intertwined nature of its double-helical structure. Type IB DNA topoisomerases are enzymes to change the topological state of DNA molecules and are essential in studying replication, transcription, and recombination of nucleic acids in vitro. NA topoisomerase II (TopoII ) is an essential chromosome-associated enzyme with activity im-plicated in the resolution of tangled DNA at cen-tromeres before anaphase onset. 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