11,283 results on '"Topoisomerase"'
Search Results
2. Genome Instability Induced by Topoisomerase Misfunction.
- Author
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Nitiss, Karin C., Bandak, Afif, Berger, James M., and Nitiss, John L.
- Subjects
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DNA topoisomerases , *DNA structure , *SMALL molecules , *DNA damage , *RIBONUCLEOTIDES , *DNA topoisomerase I , *DNA adducts - Abstract
Topoisomerases alter DNA topology by making transient DNA strand breaks (DSBs) in DNA. The DNA cleavage reaction mechanism includes the formation of a reversible protein/DNA complex that allows rapid resealing of the transient break. This mechanism allows changes in DNA topology with minimal risks of persistent DNA damage. Nonetheless, small molecules, alternate DNA structures, or mutations in topoisomerase proteins can impede the resealing of the transient breaks, leading to genome instability and potentially cell death. The consequences of high levels of enzyme/DNA adducts differ for type I and type II topoisomerases. Top1 action on DNA containing ribonucleotides leads to 2–5 nucleotide deletions in repeated sequences, while mutant Top1 enzymes can generate large deletions. By contrast, small molecules that target Top2, or mutant Top2 enzymes with elevated levels of cleavage lead to small de novo duplications. Both Top1 and Top2 have the potential to generate large rearrangements and translocations. Thus, genome instability due to topoisomerase mis-function is a potential pathogenic mechanism especially leading to oncogenic progression. Recent studies support the potential roles of topoisomerases in genetic changes in cancer cells, highlighting the need to understand how cells limit genome instability induced by topoisomerases. This review highlights recent studies that bear on these questions. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
3. Design, synthesis, and anticancer evaluation of novel N-[5-(1,3,4,5-tetrahydroxycyclohexyl)-1,3,4-thiadiazole-2-yl] benzamide analogues through integrated computational and experimental approaches
- Author
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Sujaritha Jayaraj and K. Hemalatha
- Subjects
1,3,4,5-Tetrahydroxycyclohexyl derivatives ,Molecular docking ,Topoisomerase ,SRB assay ,DMBA ,Anticancer evaluations ,Therapeutics. Pharmacology ,RM1-950 ,Pharmacy and materia medica ,RS1-441 - Abstract
Abstract Background The main aim of the current study is to develop, synthesize, in silico, in vitro and in vivo potentials of N-[5-(1,3,4,5-tetrahydroxycyclohexyl)-1,3,4-thiadiazole-2-yl] benzamide derivatives for a possible anticancer drug to improve their efficiency and selectivity against cancer cells, computational approaches aided in the rational design of these chemicals. Spectroscopic methods verified the chemical structures of the target compounds. The structures of the synthesized analogs were determined by elemental analysis, IR, 1H NMR, 13C NMR and MS. Structure shows the presence of 1,3,4, thiadiazole also responsible for anticancer activity. The 10 analogs were synthesized and showed encouraging anticancer efficacy in preliminary biological evaluation, suggesting they might be suitable lead candidates for more optimization and preclinical exploration. Result N-[5-(1,3,4,5-tetrahydroxycyclohexyl)-1,3,4-thiadiazole-2-yl] benzamide derivatives were synthesized (5a-5j) showed an optimum IC50 value in in vitro activity by SRB assay using MCF-7 as a strain, and the few selected analogs 5b,5 g & 5 h were subjected for in vivo anticancer activity by DMBA induction of tumors in mice. Conclusion Through a computational and experimental approach, this study results a way for newer derivatives for the class of anticancer drugs.
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- 2024
- Full Text
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4. Apoptotic effects of triterpenoids isolated from Pleiocarpa pycnantha leaves in cancer cells and molecular docking study of the interactions of camptothecin and ursolic acid with human caspase 3, caspase 9 and topoisomerase I.
- Author
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Akinwunmi, Olubunmi Adenike, Oso, Babatunde, Sibuyi, Nicole Remaliah Samantha, Green, Ivan, Iwuoha, Emmanuel, Meyer, Mervin, and Hussein, Ahmed
- Abstract
The aim of this study was to examine the effects of triterpenes from Pleiocarpa pycnantha leaves on the induction of apoptotic signalling in human cells. The molecular mechanisms of triterpenes isolated from P. pycnantha leaves were investigated in vitro on HeLa, MCF-7, HT-29, and KMST-6 cells. The compounds activated several markers associated with apoptosis, viz., phosphatidylserine translocation, caspase activation, oxidative stress, and topoisomerase I inhibition. Compounds 1 and 5 were non-selective, whereas compounds 2, 3, and 4 showed potential as cancer-specific agents by selectively inducing apoptosis only on cancer cells. Theoretical studies on the interactions of compound 1 with caspases −3 and −9 and topoisomerase I were carried out through a molecular docking study and illustrated that compound 1 had an equal binding affinity with the caspases and topoisomerase I comparable to that of camptothecin. The cellular pathway activated by these compounds was dependent on the compound and the cell type. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
5. Myeloperoxidase inhibition protects bone marrow mononuclear cells from DNA damage induced by the TOP2 poison anti‐cancer drug etoposide
- Author
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Ian G. Cowell and Caroline A. Austin
- Subjects
DNA damage ,etoposide ,myeloperoxidase ,myelotoxicity ,TOP2 ,topoisomerase ,Biology (General) ,QH301-705.5 - Abstract
Myeloperoxidase (MPO) is found almost exclusively in granulocytes and immature myeloid cells. It plays a key role in the innate immune system, catalysing the formation of reactive oxygen species that are important in anti‐microbial action, but MPO also oxidatively transforms the topoisomerase II (TOP2) poison etoposide to chemical forms that have elevated DNA damaging properties. TOP2 poisons such as etoposide are widely used anti‐cancer drugs, but they are linked to cases of secondary acute myeloid leukaemias through a mechanism that involves DNA damage and presumably erroneous repair leading to leukaemogenic chromosome translocations. This leads to the possibility that myeloperoxidase inhibitors could reduce the rate of therapy‐related leukaemia by protecting haematopoietic cells from TOP2 poison‐mediated genotoxic damage while preserving the anti‐cancer efficacy of the treatment. We show here that myeloperoxidase inhibition reduces etoposide‐induced TOP2B‐DNA covalent complexes and resulting DNA double‐strand break formation in primary ex vivo expanded CD34+ progenitor cells and unfractionated bone marrow mononuclear cells. Since MPO inhibitors are currently being developed as anti‐inflammatory agents this raises the possibility that repurposing of these potential new drugs could provide a means of suppressing secondary acute myeloid leukaemias associated with therapies containing TOP2 poisons.
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- 2024
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6. CX-5461 Preferentially Induces Top2α-Dependent DNA Breaks at Ribosomal DNA Loci.
- Author
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Cameron, Donald P., Sornkom, Jirawas, Alsahafi, Sameerh, Drygin, Denis, Poortinga, Gretchen, McArthur, Grant A., Hein, Nadine, Hannan, Ross, and Panov, Konstantin I.
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DNA repair ,RIBOSOMAL DNA ,RNA polymerases ,DNA damage ,GENETIC transcription - Abstract
While genotoxic chemotherapeutic agents are among the most effective tools to combat cancer, they are often associated with severe adverse effects caused by indiscriminate DNA damage in non-tumor tissue as well as increased risk of secondary carcinogenesis. This study builds on our previous work demonstrating that the RNA Polymerase I (Pol I) transcription inhibitor CX-5461 elicits a non-canonical DNA damage response and our discovery of a critical role for Topoisomerase 2α (Top2α) in the initiation of Pol I-dependent transcription. Here, we identify Top2α as a mediator of CX-5461 response in the murine Eµ-Myc B lymphoma model whereby sensitivity to CX-5461 is dependent on cellular Top2α expression/activity. Most strikingly, and in contrast to canonical Top2α poisons, we found that the Top2α-dependent DNA damage induced by CX-5461 is preferentially localized at the ribosomal DNA (rDNA) promoter region, thereby highlighting CX-5461 as a loci-specific DNA damaging agent. This mechanism underpins the efficacy of CX-5461 against certain types of cancer and can be used to develop effective non-genotoxic anticancer drugs. [ABSTRACT FROM AUTHOR]
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- 2024
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7. Discovery of new Schiff bases of the disalicylic acid scaffold as DNA gyrase and topoisomerase IV inhibitors endowed with antibacterial properties.
- Author
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Al-Wahaibi, Lamya H., Mahmoud, Mohamed A., Alzahrani, Hayat Ali, Abou-Zied, Hesham A., Gomaa, Hesham A. M., Youssif, Bahaa G. M., Brase, Stefan, Rabea, Safwat M., Amorim, Juliana, Baki, Vijaya Bhaskar, and Wu, Chunli
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DNA topoisomerase II , *SCHIFF bases , *DNA topoisomerases , *ANTIBACTERIAL agents , *SALICYLIC acid - Abstract
DNA gyrase and topoisomerase IV show great potential as targets for antibacterial medicines. In recent decades, various categories of small molecule inhibitors have been identified; however, none have been effective in the market. For the first time, we developed a series of disalicylic acid methylene/Schiff bases hybrids (5a-k) to act as antibacterial agents targeting DNA gyrase and topoisomerase IV. The findings indicated that the new targets 5f-k exhibited significant antibacterial activity against Gram-positive and Gram-negative bacteria, with efficacy ranging from 75% to 115% of the standard ciprofloxacin levels. Compound 5h demonstrated the greatest efficacy compared to the other compounds tested, with minimum inhibitory concentration (MIC) values of 0.030, 0.065, and 0.060 ^ig/mL against S. aureus, E. coli, and P. aeruginosa. 5h had a MIC value of 0.050 ^ig/mL against B. subtilis, which is five times less potent than ciprofloxacin. The inhibitory efficacy of the most potent antibacterial derivatives 5f, 5h, 5i, and 5k against E. coli DNA gyrase was assessed. The tested compounds demonstrated inhibitory effects on E. coli DNA gyrase, with IC50 values ranging from 92 to 112 nM. These results indicate that 5f, 5h, 5i, and 5k are more effective than the reference novobiocin, which had an IC50 value of 170 nM. Compounds 5f, 5h, 5i, and 5k were subjected to additional assessment against E. coli topoisomerase IV. Compounds 5h and 5i, which have the highest efficacy in inhibiting E. coli gyrase, also demonstrated promising effects on topoisomerase IV. Compounds 5h and 5i exhibit IC50 values of 3.50 ^M and 5.80 ^M, respectively. These results are much lower and more potent than novobiocin's IC50 value of 11 ^M. Docking studies demonstrate the potential of compound 5h as an effective dual inhibitor against E. coli DNA gyrase and topoisomerase IV, with ADMET analysis indicating promising pharmacokinetic profiles for antibacterial drug development. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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8. IN SILICO DESIGNING AND SCREENING OF CARBAZOLE DERIVATIVES AS TOPOISOMERASE II INHIBITORS.
- Author
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Jose, Elseena, Francis, Prasanth, Joy, Mariya, Joseph, Shilpa, Sukumaran, Sunitha, and Baby, Sini
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DNA topoisomerase II , *DNA topoisomerase I , *MOLECULAR docking , *CARBAZOLE derivatives , *MOLECULAR dynamics - Abstract
Due to side effects and drug resistance, cancer treatment choices are limited despite continued attempts. Topoisomerases are essential for several cellular processes and human topoisomerase I and topoisomerase II alpha inhibitors have proven to be an effective chemotherapeutic alternative for a wide range of cancers, due to its fast proliferating cells and the higher level of these enzymes in solid tumors relative to normal tissue. In this study, around 30 carbazole derivatives were designed using Chem Draw Ultra and their ability to inhibit topoisomerase II was investigated using in silico docking studies and molecular dynamics. The results revealed ligand 10 and ligand 25 held good binding energy scores of -9.21 and -9.50, Kcal mol-1 respectively, with good interaction. ADMET analysis assured the drug likeness and better GI absorption than the parent drug and dexrazoxane. Desmond module of Schrödinger Suite was used and it demonstrated the stability in ligand receptor complexes. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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9. Myeloperoxidase inhibition protects bone marrow mononuclear cells from DNA damage induced by the TOP2 poison anti‐cancer drug etoposide.
- Author
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Cowell, Ian G. and Austin, Caroline A.
- Subjects
BONE marrow cells ,DOUBLE-strand DNA breaks ,DNA damage ,MYELOPEROXIDASE ,DNA repair ,ANTINEOPLASTIC agents ,DRUG toxicity ,DNA topoisomerase I - Abstract
Myeloperoxidase (MPO) is found almost exclusively in granulocytes and immature myeloid cells. It plays a key role in the innate immune system, catalysing the formation of reactive oxygen species that are important in anti‐microbial action, but MPO also oxidatively transforms the topoisomerase II (TOP2) poison etoposide to chemical forms that have elevated DNA damaging properties. TOP2 poisons such as etoposide are widely used anti‐cancer drugs, but they are linked to cases of secondary acute myeloid leukaemias through a mechanism that involves DNA damage and presumably erroneous repair leading to leukaemogenic chromosome translocations. This leads to the possibility that myeloperoxidase inhibitors could reduce the rate of therapy‐related leukaemia by protecting haematopoietic cells from TOP2 poison‐mediated genotoxic damage while preserving the anti‐cancer efficacy of the treatment. We show here that myeloperoxidase inhibition reduces etoposide‐induced TOP2B‐DNA covalent complexes and resulting DNA double‐strand break formation in primary ex vivo expanded CD34+ progenitor cells and unfractionated bone marrow mononuclear cells. Since MPO inhibitors are currently being developed as anti‐inflammatory agents this raises the possibility that repurposing of these potential new drugs could provide a means of suppressing secondary acute myeloid leukaemias associated with therapies containing TOP2 poisons. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
10. Tumor growth-arrest effect of tetrahydroquinazoline-derivative human topoisomerase II-alpha inhibitor in HPV-negative head and neck squamous cell carcinoma
- Author
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Patrizia Sarogni, Nicoletta Brindani, Agata Zamborlin, Alessandra Gonnelli, Michele Menicagli, Ana Katrina Mapanao, Federico Munafò, Marco De Vivo, and Valerio Voliani
- Subjects
Topoisomerase ,Head and neck ,Alternative models ,Tetrahydroquinazoline ,Medicine ,Science - Abstract
Abstract Oral malignancies continue to have severe morbidity with less than 50% long-term survival despite the advancement in the available therapies. There is a persisting demand for new approaches to establish more efficient strategies for their treatment. In this regard, the human topoisomerase II (topoII) enzyme is a validated chemotherapeutics target, as topoII regulates vital cellular processes such as DNA replication, transcription, recombination, and chromosome segregation in cells. TopoII inhibitors are currently used to treat some neoplasms such as breast and small cells lung carcinomas. Additionally, topoII inhibitors are under investigation for the treatment of other cancer types, including oral cancer. Here, we report the therapeutic effect of a tetrahydroquinazoline derivative (named ARN21934) that preferentially inhibits the alpha isoform of human topoII. The treatment efficacy of ARN21934 has been evaluated in 2D cell cultures, 3D in vitro systems, and in chick chorioallantoic membrane cancer models. Overall, this work paves the way for further preclinical developments of ARN21934 and possibly other topoII alpha inhibitors of this promising chemical class as a new chemotherapeutic approach for the treatment of oral neoplasms.
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- 2024
- Full Text
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11. Palindromic carbazole derivatives: unveiling their antiproliferative effect via topoisomerase II catalytic inhibition and apoptosis induction
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Mateusz Olszewski, Natalia Maciejewska, Anoop Kallingal, Agnieszka Chylewska, Aleksandra M. Dąbrowska, Małgorzata Biedulska, Mariusz Makowski, José M. Padrón, and Maciej Baginski
- Subjects
Apoptosis ,cancer ,carbazole ,topoisomerase ,Therapeutics. Pharmacology ,RM1-950 - Abstract
AbstractHuman DNA topoisomerases are essential for crucial cellular processes, including DNA replication, transcription, chromatin condensation, and maintenance of its structure. One of the significant strategies employed in cancer treatment involves the inhibition of a specific type of topoisomerase, known as topoisomerase II (Topo II). Carbazole derivatives, recognised for their varied biological activities, have recently become a significant focus in oncological research. This study assesses the efficacy of three symmetrically substituted carbazole derivatives: 2,7-Di(2-furyl)-9H-carbazole (27a), 3,6-Di(2-furyl)-9H-carbazole (36a), and 3,6-Di(2-thienyl)-9H-carbazole (36b) – as anticancer agents. Among investigated carbazole derivatives, compound 3,6-di(2-furyl)-9H-carbazole bearing two furan moieties emerged as a novel catalytic inhibitor of Topo II. Notably, 3,6-di(2-furyl)-9H-carbazole effectively selectively inhibited the relaxation and decatenation activities of Topo IIα, with minimal effects on the IIβ isoform. These findings underscore the potential of compound 3,6-Di(2-furyl)-9H-carbazole as a promising lead candidate warranting further investigation in the realm of anticancer drug development.
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- 2024
- Full Text
- View/download PDF
12. Tumor growth-arrest effect of tetrahydroquinazoline-derivative human topoisomerase II-alpha inhibitor in HPV-negative head and neck squamous cell carcinoma.
- Author
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Sarogni, Patrizia, Brindani, Nicoletta, Zamborlin, Agata, Gonnelli, Alessandra, Menicagli, Michele, Mapanao, Ana Katrina, Munafò, Federico, De Vivo, Marco, and Voliani, Valerio
- Subjects
- *
BREAST , *SQUAMOUS cell carcinoma , *NECK , *DNA topoisomerase I , *HUMAN papillomavirus , *DNA topoisomerase II , *CHORIOALLANTOIS , *SMALL cell carcinoma - Abstract
Oral malignancies continue to have severe morbidity with less than 50% long-term survival despite the advancement in the available therapies. There is a persisting demand for new approaches to establish more efficient strategies for their treatment. In this regard, the human topoisomerase II (topoII) enzyme is a validated chemotherapeutics target, as topoII regulates vital cellular processes such as DNA replication, transcription, recombination, and chromosome segregation in cells. TopoII inhibitors are currently used to treat some neoplasms such as breast and small cells lung carcinomas. Additionally, topoII inhibitors are under investigation for the treatment of other cancer types, including oral cancer. Here, we report the therapeutic effect of a tetrahydroquinazoline derivative (named ARN21934) that preferentially inhibits the alpha isoform of human topoII. The treatment efficacy of ARN21934 has been evaluated in 2D cell cultures, 3D in vitro systems, and in chick chorioallantoic membrane cancer models. Overall, this work paves the way for further preclinical developments of ARN21934 and possibly other topoII alpha inhibitors of this promising chemical class as a new chemotherapeutic approach for the treatment of oral neoplasms. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
13. Variation of Structure and Cellular Functions of Type IA Topoisomerases across the Tree of Life.
- Author
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Tan, Kemin and Tse-Dinh, Yuk-Ching
- Subjects
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DNA topoisomerase I , *SINGLE-strand DNA breaks , *CELL physiology , *CELL anatomy , *NUCLEIC acids , *PROTEIN domains - Abstract
Topoisomerases regulate the topological state of cellular genomes to prevent impediments to vital cellular processes, including replication and transcription from suboptimal supercoiling of double-stranded DNA, and to untangle topological barriers generated as replication or recombination intermediates. The subfamily of type IA topoisomerases are the only topoisomerases that can alter the interlinking of both DNA and RNA. In this article, we provide a review of the mechanisms by which four highly conserved N-terminal protein domains fold into a toroidal structure, enabling cleavage and religation of a single strand of DNA or RNA. We also explore how these conserved domains can be combined with numerous non-conserved protein sequences located in the C-terminal domains to form a diverse range of type IA topoisomerases in Archaea, Bacteria, and Eukarya. There is at least one type IA topoisomerase present in nearly every free-living organism. The variation in C-terminal domain sequences and interacting partners such as helicases enable type IA topoisomerases to conduct important cellular functions that require the passage of nucleic acids through the break of a single-strand DNA or RNA that is held by the conserved N-terminal toroidal domains. In addition, this review will exam a range of human genetic disorders that have been linked to the malfunction of type IA topoisomerase. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
14. Ruthenium(II) complex with 2-mercaptothiazoline ligand induces selective cytotoxicity involving DNA damage and apoptosis in melanoma cells.
- Author
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de Melo, Matheus Reis Santos, Ribeiro, Arthur Barcelos, Fernandes, Gabriela, Squarisi, Iara Silva, de Melo Junqueira, Marcela, Batista, Alzir Azevedo, da Silva, Monize Martins, and Tavares, Denise Crispim
- Subjects
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DACARBAZINE , *CYTOTOXINS , *DNA damage , *LIGANDS (Biochemistry) , *MELANOMA , *RUTHENIUM , *CELL morphology - Abstract
Melanoma is the most aggressive and lethal type of skin cancer due to its characteristics such as high metastatic potential and low response rate to existing treatment modalities. In this way, new drug prototypes are being studied to solve the problem of treating patients with melanoma. Among these, ruthenium-based metallopharmaceuticals may be promising alternatives due to their antitumor characteristics and low systemic toxicity. In this context, the present study evaluated the antineoplastic effect of the ruthenium complex [Ru(mtz)(dppe)2]PF6-2-mercaptothiazoline-di-1,2-bis(diphenylphosphine) ethaneruthenium(II), namely RuMTZ, on human melanoma (A-375) and murine (B16-F10) cells, considering different approaches. Through XTT colorimetric and clonogenic efficiency assays, the complex revealed the selective cytotoxic activity, with the lowest IC50 (0.4 µM) observed for A375 cells. RuMTZ also induced changes in cell morphology, increased cell population in the sub-G0 phase and inhibiting cell migration. The levels of γH2AX and cleaved caspase 3 proteins were increased in both cell lines treated with RuMTZ. These findings indicated that the cytotoxic activity of RuMTZ on melanoma cells is related, at least in part, to the induction of DNA damage and apoptosis. Therefore, RuMTZ exhibited promising antineoplastic activity against melanoma cells. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
15. Rapid, DNA-induced interface swapping by DNA gyrase
- Author
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Thomas RM Germe, Natassja G Bush, Victoria M Baskerville, Dominik Saman, Justin LP Benesch, and Anthony Maxwell
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topoisomerase ,antibacterials ,enzymology ,genome dynamics ,DNA gyrase ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
DNA gyrase, a ubiquitous bacterial enzyme, is a type IIA topoisomerase formed by heterotetramerisation of 2 GyrA subunits and 2 GyrB subunits, to form the active complex. DNA gyrase can loop DNA around the C-terminal domains (CTDs) of GyrA and pass one DNA duplex through a transient double-strand break (DSB) established in another duplex. This results in the conversion from a positive (+1) to a negative (–1) supercoil, thereby introducing negative supercoiling into the bacterial genome by steps of 2, an activity essential for DNA replication and transcription. The strong protein interface in the GyrA dimer must be broken to allow passage of the transported DNA segment and it is generally assumed that the interface is usually stable and only opens when DNA is transported, to prevent the introduction of deleterious DSBs in the genome. In this paper, we show that DNA gyrase can exchange its DNA-cleaving interfaces between two active heterotetramers. This so-called interface ‘swapping’ (IS) can occur within a few minutes in solution. We also show that bending of DNA by gyrase is essential for cleavage but not for DNA binding per se and favors IS. Interface swapping is also favored by DNA wrapping and an excess of GyrB. We suggest that proximity, promoted by GyrB oligomerization and binding and wrapping along a length of DNA, between two heterotetramers favors rapid interface swapping. This swapping does not require ATP, occurs in the presence of fluoroquinolones, and raises the possibility of non-homologous recombination solely through gyrase activity. The ability of gyrase to undergo interface swapping explains how gyrase heterodimers, containing a single active-site tyrosine, can carry out double-strand passage reactions and therefore suggests an alternative explanation to the recently proposed ‘swivelling’ mechanism for DNA gyrase (Gubaev et al., 2016).
- Published
- 2024
- Full Text
- View/download PDF
16. Effect of Metal-Binding Antimicrobial Peptide SIF4 on Topoisomerases Activity and Intracellular Nucleic Acid Biosynthesis in Escherichia coli
- Author
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LI Yuzhen, XIAO Huaiqiu, ZHOU Huiheng, LI Lan, KUANG Yan, LIU Miao, ZHAO Mouming
- Subjects
metal-binding antimicrobial peptide ,escherichia coli ,topoisomerase ,nucleic acid ,antimicrobial agent ,Food processing and manufacture ,TP368-456 - Abstract
To systematically elucidate how metal-binding antimicrobial peptide SIF4 exerts its antimicrobial activity by targeting DNA topoisomerase without destroying the cytoplasmic membrane, Escherichia coli was used as a model strain to investigate the binding mode of SIF4 with its genomic DNA and the impact of SIF4 on DNA topoisomerase I and II activities and intracellular nucleic acid biosynthesis. Results showed that SIF4 could bind to genomic DNA in a manner similar to ethidium bromide (EB) intercalation with strong inhibitory effect on topoisomerase I but weak effect on topoisomerase II, and catalyzed RNA transcription to exert antimicrobial activity by interfering with the unwinding of negative DNA supercoils and RNA polymerase binding. It was also found that the biosynthesis of intracellular DNA and RNA was inhibited to different degrees after 12 h treatment with SIF4, which exhibited a good dose-effect relationship. There was no significant difference in the amounts of intracellular DNA and RNA between the 1/2 minimum inhibitory concentration (MIC) group and the control group (P > 0.05), but there was a significant difference between the MIC and 2 MIC groups and the control group (P < 0.05). Our results may provide theoretical support for the application of SIF4 in the biocontrol of foodborne E. coli.
- Published
- 2024
- Full Text
- View/download PDF
17. Genome Instability Induced by Topoisomerase Misfunction
- Author
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Karin C. Nitiss, Afif Bandak, James M. Berger, and John L. Nitiss
- Subjects
topoisomerase ,mutation ,genome instability ,Top1 ,Top2 ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Topoisomerases alter DNA topology by making transient DNA strand breaks (DSBs) in DNA. The DNA cleavage reaction mechanism includes the formation of a reversible protein/DNA complex that allows rapid resealing of the transient break. This mechanism allows changes in DNA topology with minimal risks of persistent DNA damage. Nonetheless, small molecules, alternate DNA structures, or mutations in topoisomerase proteins can impede the resealing of the transient breaks, leading to genome instability and potentially cell death. The consequences of high levels of enzyme/DNA adducts differ for type I and type II topoisomerases. Top1 action on DNA containing ribonucleotides leads to 2–5 nucleotide deletions in repeated sequences, while mutant Top1 enzymes can generate large deletions. By contrast, small molecules that target Top2, or mutant Top2 enzymes with elevated levels of cleavage lead to small de novo duplications. Both Top1 and Top2 have the potential to generate large rearrangements and translocations. Thus, genome instability due to topoisomerase mis-function is a potential pathogenic mechanism especially leading to oncogenic progression. Recent studies support the potential roles of topoisomerases in genetic changes in cancer cells, highlighting the need to understand how cells limit genome instability induced by topoisomerases. This review highlights recent studies that bear on these questions.
- Published
- 2024
- Full Text
- View/download PDF
18. Disease-associated mutations in topoisomerase IIβ result in defective NK cells.
- Author
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Broderick, Lori, Clay, Gwendolyn M, Blum, Robert H, Liu, Yang, McVicar, Rachael, Papes, Fabio, Booshehri, Laela M, Cowell, Ian G, Austin, Caroline A, Putnam, Christopher D, and Kaufman, Dan S
- Subjects
Killer Cells ,Natural ,Cell Line ,Animals ,Humans ,Mice ,Craniofacial Abnormalities ,Limb Deformities ,Congenital ,Urogenital Abnormalities ,Mutation ,Induced Pluripotent Stem Cells ,Primary Immunodeficiency Diseases ,Immunodeficiency ,iPSC ,natural killer cells ,topoisomerase ,Clinical Research ,Stem Cell Research - Induced Pluripotent Stem Cell - Human ,Rare Diseases ,Stem Cell Research ,Stem Cell Research - Induced Pluripotent Stem Cell ,Pediatric ,Genetics ,2.1 Biological and endogenous factors ,Aetiology ,Immunology ,Allergy - Abstract
BackgroundHoffman syndrome is a syndromic, inborn error of immunity due to autosomal-dominant mutations in TOP2B, an essential gene required to alleviate topological stress during DNA replication and gene transcription. Although mutations identified in patients lead to a block in B-cell development and the absence of circulating B cells, an effect on natural killer (NK) cells was not previously examined.ObjectiveWe sought to determine whether disease-associated mutations in TOP2B impact NK-cell development and function.MethodsUsing a knockin murine model and patient-derived induced pluripotent stem cells (iPSCs), we investigated NK-cell development in mouse bone marrow and spleen, and performed immunophenotyping by flow cytometry, gene expression, and functional assessment of cytotoxic activity in murine NK cells, and human IPSC-derived NK cells.ResultsMature NK cells were reduced in the periphery of TOP2B knockin mice consistent with patient reports, with reduced cytotoxicity toward target cell lines. IPSCs were successfully derived from patients with Hoffman syndrome, but under optimal conditions showed reduced cytotoxicity compared with iPSC-derived NK cells from healthy controls.ConclusionsHoffman syndrome-associated mutations in TOP2B impact NK-cell development and function in murine and human models.
- Published
- 2022
19. Structure of Escherichia coli exonuclease VII.
- Author
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Chuan Liu, Hauk, Glenn, Qianyun Yan, and Berger, James M.
- Subjects
- *
ESCHERICHIA coli , *DOUBLE-strand DNA breaks , *CATALYTIC domains , *SINGLE-stranded DNA , *DNA repair - Abstract
Exonuclease VII (ExoVII) is a ubiquitous bacterial nuclease. Encoded by the xseA and xseB genes, ExoVII participates in multiple nucleic acid-dependent pathways including the processing of multicopy single-stranded DNA and the repair of covalent DNA-protein crosslinks (DPCs). Although many biochemical properties of ExoVII have been defined, little is known about its structure/function relationships. Here, we use cryoelectron microscopy (cryoEM) to determine that Escherichia coli ExoVII comprises a highly elongated XseA4·XseB24 holo-complex. Each XseA subunit dimerizes through a central extended a-helical segment decorated by six XseB subunits and a C-terminal, domain-swapped ß-barrel element; two XseA2·XseB12 subcomplexes further associate using N-terminal OB (oligonucleotide/oligosaccharide-binding) folds and catalytic domains to form a spindle-shaped, catenated octaicosamer. The catalytic domains of XseA, which adopt a nuclease fold related to 3-dehydroquinate dehydratases, are sequestered in the center of the complex and accessible only through large pores formed between XseA tetramers. The architectural organization of ExoVII, combined with biochemical studies, indicate that substrate selectivity is controlled by steric access to its nuclease elements and that tetramer dissociation results from substrate DNA binding. Despite a lack of sequence and fold homology, the physical organization of ExoVII is reminiscent of Mre11·Rad50/SbcCD ATP (adenosine triphosphate)-dependent nucleases used in the repair of double-stranded DNA breaks, including those formed by DPCs through aberrant topoisomerase activity, suggesting that there may have been convergent evolutionary pressure to contend with such damage events. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
20. 金属抗菌肽 SIF4对大肠杆菌拓扑异构酶活性及 胞内核酸合成的影响.
- Author
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李玉珍, 肖怀秋, 周慧恒, 李 篮, 匡 燕, 刘 淼, and 赵谋明
- Subjects
ANTIMICROBIAL peptides ,NUCLEIC acids ,ESCHERICHIA coli ,ANTI-infective agents - Abstract
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- 2024
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21. The involvement of hepatic cytochrome P450s in the cytotoxicity of lapatinib.
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Chen, Si, Li, Xilin, Li, Yuxi, He, Xiaobo, Bryant, Matthew, Qin, Xuan, Li, Feng, Seo, Ji-Eun, Guo, Xiaoqing, Mei, Nan, and Guo, Lei
- Subjects
- *
CYTOTOXINS , *LAPATINIB , *HER2 positive breast cancer , *PROTEIN-tyrosine kinase inhibitors , *LIVER cells , *DNA damage , *CYTOCHROME P-450 , *CYTOCHROME c - Abstract
Lapatinib, an oral tyrosine kinase inhibitor used as a first-line treatment for HER2-positive breast cancer, has been reported to be associated with hepatotoxicity; however, the underlying mechanisms remain unclear. In this study, we report that lapatinib causes cytotoxicity in multiple types of hepatic cells, including primary human hepatocytes, HepaRG cells, and HepG2 cells. A 24-h treatment with lapatinib induced cell cycle disturbances, apoptosis, and DNA damage, and decreased the protein levels of topoisomerase in HepG2 cells. We investigated the role of cytochrome P450 (CYP)-mediated metabolism in lapatinib-induced cytotoxicity using our previously established HepG2 cell lines, which express each of 14 CYPs (1A1, 1A2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 3A4, 3A5, and 3A7). We demonstrate that lapatinib is metabolized by CYP1A1, 3A4, 3A5, and 3A7. Among these, lapatinib-induced cytotoxicity and DNA damage were attenuated in cells overexpressing CYP3A5 or 3A7. Additionally, we measured the production of three primary metabolites of lapatinib (O -dealkylated lapatinib, N -dealkylated lapatinib, and N -hydroxy lapatinib) in CYP1A1-, 3A4-, 3A5-, and 3A7-overexpressing HepG2 cells. We compared the cytotoxicity of lapatinib and its 3 metabolites in primary human hepatocytes, HepaRG cells, and HepG2 cells and demonstrated that N -dealkylated lapatinib is more toxic than the parent drug and the other metabolites. Taken together, our results indicate that lapatinib-induced cytotoxicity involves multiple mechanisms, such as apoptosis and DNA damage; that N -dealkylated lapatinib is a toxic metabolite contributing to the toxic effect of lapatinib; and that CYP3A5- and 3A7-mediated metabolism plays a role in attenuating the cytotoxicity of lapatinib. [ABSTRACT FROM AUTHOR]
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- 2024
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22. New Imidazo[4,5-α] Acridine: Synthesis and Studying the Molecular Dynamics Simulation of Its Interaction with the Topoisomerase Enzyme.
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Toosi, Maryam Rangamiz, Pordel, Mehdi, and Bozorgmehr, Mohammad Reza
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- *
MOLECULAR dynamics , *DNA topoisomerase I , *ACRIDINE derivatives , *ACRIDINE , *DENSITY functionals , *DENSITY functional theory , *HYDROGEN bonding interactions - Abstract
Acridine and imidazole were combined and synthesized 3,8-disubstituted-propyl-3H-imidazo[4,5-α]acridine-11-carbonitrile, as a new derivative. The interaction of this new compound with the topoisomerase enzyme was studied by molecular dynamics simulation. The 3,8-disubstituted-3H-imidazo[4,5-α]acridine-11-carbonitrile structure has been optimized by the density functional theory method. According to the results obtained from the molecular dynamics simulation, Arg364, Lys532, Asp533, Tyr537, Arg590, Cys630, Asn631, Gln633 and Adenine11 interact with the ligand by hydrophobic interactions and Arg488 and Adenine12 interact with the ligand by hydrogen bond interactions. Due to the fact that some of these residues, Arg488 and Arg590 are located in the enzyme active site, the new ligand appears to be inhibitory effect. Also, the calculation of the Harmonic Oscillator Model for Aromaticity (HOMA) index showed that the 5-membered ring of ligand and the 6-membered ring attached to the 5-membered ring had more reactivity with the enzyme. The contribution of charged residues in the binding free energy of the ligand is greater than the uncharged residues. [ABSTRACT FROM AUTHOR]
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- 2024
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23. The roles of nucleoid-associated proteins and topoisomerases in chromosome structure, strand segregation, and the generation of phenotypic heterogeneity in bacteria.
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Norris, Vic, Kayser, Clara, Muskhelishvili, Georgi, and Konto-Ghiorghi, Yoan
- Subjects
- *
CHROMOSOME structure , *BACTERIAL chromosomes , *DNA topoisomerase II , *PHENOTYPES , *CARRIER proteins , *CHROMOSOMES - Abstract
How to adapt to a changing environment is a fundamental, recurrent problem confronting cells. One solution is for cells to organize their constituents into a limited number of spatially extended, functionally relevant, macromolecular assemblies or hyperstructures, and then to segregate these hyperstructures asymmetrically into daughter cells. This asymmetric segregation becomes a particularly powerful way of generating a coherent phenotypic diversity when the segregation of certain hyperstructures is with only one of the parental DNA strands and when this pattern of segregation continues over successive generations. Candidate hyperstructures for such asymmetric segregation in prokaryotes include those containing the nucleoid-associated proteins (NAPs) and the topoisomerases. Another solution to the problem of creating a coherent phenotypic diversity is by creating a growth-environment-dependent gradient of supercoiling generated along the replication origin-to-terminus axis of the bacterial chromosome. This gradient is modulated by transcription, NAPs, and topoisomerases. Here, we focus primarily on two topoisomerases, TopoIV and DNA gyrase in Escherichia coli , on three of its NAPs (H-NS, HU, and IHF), and on the single-stranded binding protein, SSB. We propose that the combination of supercoiling-gradient-dependent and strand-segregation-dependent topoisomerase activities result in significant differences in the supercoiling of daughter chromosomes, and hence in the phenotypes of daughter cells. [ABSTRACT FROM AUTHOR]
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- 2023
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24. Structure-function analysis of the ATPase domain of African swine fever virus topoisomerase
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Wenhua Kuang, Yan Zhao, Jinyue Li, and Zengqin Deng
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African swine fever virus ,topoisomerase ,ATPase domain ,crystal structure ,Microbiology ,QR1-502 - Abstract
ABSTRACTType II topoisomerase utilizes the energy from ATP hydrolysis to alter DNA topology during genome replication and transcription. The ATPase domain of this enzyme is required for ATP hydrolysis and plays a crucial role in coupling DNA binding and ATP turnover with the DNA strand passage reaction. The African swine fever virus (ASFV) specifically encodes a topoisomerase II (topo II), which is critical for viral replication and an attractive target for antiviral development. Here, we present a high-resolution crystal structure of the ASFV topo II ATPase domain complexed with the substrate analog AMPPNP. Structural comparison reveals that the ASFV topo II ATPase domain shares a conserved overall structure with its homologs from eukaryotes and prokaryotes but also has three characteristic regions, including the intra-molecular interface formed by the ATP-lid and QTK loop as well as helix α9, the K-loop in the transducer domain, and the antennae-like α-helix at the ATP binding domain. Mutating the key residues within these three regions impairs or abolishes the basal and DNA-stimulated ATPase activities and reduces or eliminates the relaxation activity of the holoenzyme. Our data indicate that all three regions are functionally important for the ATPase and relaxation activities and strongly suggest that ATP hydrolysis, DNA binding, and strand passage are highly coupled and managed by the allosteric coordination of multiple domains of the type II topoisomerase. Moreover, we find a promising druggable pocket in the dimeric interface of the ASFV topo II ATPase domain, which will benefit future anti-ASFV drug development.IMPORTANCEThe ATPase domain of type II topoisomerase provides energy by hydrolyzing ATP and coordinates with the DNA-binding/cleavage domain to drive and control DNA transport. The precise molecular mechanisms of how these domains respond to DNA binding and ATP hydrolysis signals and communicate with each other remain elusive. We determine the first high-resolution crystal structure of the ATPase domain of African swine fever virus (ASFV) topo II in complex with AMPPNP and biochemically investigate its function in ATPase and DNA relaxation activities. Importantly, we find that mutations at three characteristic regions of the ASFV ATPase domain produce parallel effects on the basal/DNA-stimulated ATPase and relaxation activities, implying the tight coupling of the ATP hydrolysis and strand passage process. Therefore, our data provide important implications for understanding the strand passage mechanism of the type II topoisomerase and the structural basis for developing ATPase domain-targeting antivirals against ASFV.
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- 2024
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25. Narciclasine, a novel topoisomerase I inhibitor, exhibited potent anti-cancer activity against cancer cells
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Meichen Wang, Leilei Liang, Rong Wang, Shutao Jia, Chang Xu, Yuting Wang, Min Luo, Qiqi Lin, Min Yang, Hongyu Zhou, Dandan Liu, and Chen Qing
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Topoisomerase ,Narciclasine (NCS) ,Topo I-DNA covalent complex ,DNA damage ,Cell cycle ,Apoptosis ,Botany ,QK1-989 - Abstract
Abstract DNA topoisomerases are essential nuclear enzymes in correcting topological DNA errors and maintaining DNA integrity. Topoisomerase inhibitors are a significant class of cancer chemotherapeutics with a definite curative effect. Natural products are a rich source of lead compounds for drug discovery, including anti-tumor drugs. In this study, we found that narciclasine (NCS), an amaryllidaceae alkaloid, is a novel inhibitor of topoisomerase I (topo I). Our data demonstrated that NCS inhibited topo I activity and reversed its unwinding effect on p-HOT DNA substrate. However, it had no obvious effect on topo II activity. The molecular mechanism of NCS inhibited topo I showed that NCS did not stabilize topo-DNA covalent complexes in cells, indicating that NCS is not a topo I poison. A blind docking result showed that NCS could bind to topo I, suggesting that NCS might be a topo I suppressor. Additionally, NCS exhibited a potent anti-proliferation effect in various cancer cells. NCS arrested the cell cycle at G2/M phase and induced cell apoptosis. Our study reveals the antitumor mechanisms of NCS and provides a good foundation for the development of anti-cancer drugs based on topo I inhibition. Graphical abstract
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- 2023
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26. CX-5461 Preferentially Induces Top2α-Dependent DNA Breaks at Ribosomal DNA Loci
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Donald P. Cameron, Jirawas Sornkom, Sameerh Alsahafi, Denis Drygin, Gretchen Poortinga, Grant A. McArthur, Nadine Hein, Ross Hannan, and Konstantin I. Panov
- Subjects
ribosome biogenesis ,topoisomerase ,DNA damage pathway ,double-strand breaks ,RNA Polymerase I ,nucleolar surveillance pathway ,Biology (General) ,QH301-705.5 - Abstract
While genotoxic chemotherapeutic agents are among the most effective tools to combat cancer, they are often associated with severe adverse effects caused by indiscriminate DNA damage in non-tumor tissue as well as increased risk of secondary carcinogenesis. This study builds on our previous work demonstrating that the RNA Polymerase I (Pol I) transcription inhibitor CX-5461 elicits a non-canonical DNA damage response and our discovery of a critical role for Topoisomerase 2α (Top2α) in the initiation of Pol I-dependent transcription. Here, we identify Top2α as a mediator of CX-5461 response in the murine Eµ-Myc B lymphoma model whereby sensitivity to CX-5461 is dependent on cellular Top2α expression/activity. Most strikingly, and in contrast to canonical Top2α poisons, we found that the Top2α-dependent DNA damage induced by CX-5461 is preferentially localized at the ribosomal DNA (rDNA) promoter region, thereby highlighting CX-5461 as a loci-specific DNA damaging agent. This mechanism underpins the efficacy of CX-5461 against certain types of cancer and can be used to develop effective non-genotoxic anticancer drugs.
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- 2024
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27. Direct interaction of DNA repair protein tyrosyl DNA phosphodiesterase 1 and the DNA ligase III catalytic domain is regulated by phosphorylation of its flexible N-terminus.
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Rashid, Ishtiaque, Hammel, Michal, Sverzhinsky, Aleksandr, Tsai, Miaw-Sheue, Pascal, John M, Tainer, John A, and Tomkinson, Alan E
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DNA ligation ,electron microscopy ,phosphorylation ,protein structure ,small angle x-ray scattering ,topoisomerase ,Genetics ,Cancer ,1.1 Normal biological development and functioning ,2.1 Biological and endogenous factors ,Generic health relevance ,Biochemistry & Molecular Biology ,Chemical Sciences ,Biological Sciences ,Medical and Health Sciences - Abstract
Tyrosyl DNA phosphodiesterase 1 (TDP1) and DNA Ligase IIIα (LigIIIα) are key enzymes in single-strand break (SSB) repair. TDP1 removes 3'-tyrosine residues remaining after degradation of DNA topoisomerase (TOP) 1 cleavage complexes trapped by either DNA lesions or TOP1 inhibitors. It is not known how TDP1 is linked to subsequent processing and LigIIIα-catalyzed joining of the SSB. Here we define a direct interaction between the TDP1 catalytic domain and the LigIII DNA-binding domain (DBD) regulated by conformational changes in the unstructured TDP1 N-terminal region induced by phosphorylation and/or alterations in amino acid sequence. Full-length and N-terminally truncated TDP1 are more effective at correcting SSB repair defects in TDP1 null cells compared with full-length TDP1 with amino acid substitutions of an N-terminal serine residue phosphorylated in response to DNA damage. TDP1 forms a stable complex with LigIII170-755, as well as full-length LigIIIα alone or in complex with the DNA repair scaffold protein XRCC1. Small-angle X-ray scattering and negative stain electron microscopy combined with mapping of the interacting regions identified a TDP1/LigIIIα compact dimer of heterodimers in which the two LigIII catalytic cores are positioned in the center, whereas the two TDP1 molecules are located at the edges of the core complex flanked by highly flexible regions that can interact with other repair proteins and SSBs. As TDP1and LigIIIα together repair adducts caused by TOP1 cancer chemotherapy inhibitors, the defined interaction architecture and regulation of this enzyme complex provide insights into a key repair pathway in nonmalignant and cancer cells.
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- 2021
28. Prospects of Topoisomerase Inhibitors as Promising Anti-Cancer Agents.
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Yakkala, Prasanna Anjaneyulu, Penumallu, Naveen Reddy, Shafi, Syed, and Kamal, Ahmed
- Subjects
- *
DNA topoisomerase I , *ANTINEOPLASTIC agents , *DOUBLE-strand DNA breaks , *DNA topoisomerase II , *DNA replication , *METASTASIS , *MULTIDRUG resistance - Abstract
Topoisomerases are very important enzymes that regulate DNA topology and are vital for biological actions like DNA replication, transcription, and repair. The emergence and spread of cancer has been intimately associated with topoisomerase dysregulation. Topoisomerase inhibitors have consequently become potential anti-cancer medications because of their ability to obstruct the normal function of these enzymes, which leads to DNA damage and subsequently causes cell death. This review emphasizes the importance of topoisomerase inhibitors as marketed, clinical and preclinical anti-cancer medications. In the present review, various types of topoisomerase inhibitors and their mechanisms of action have been discussed. Topoisomerase I inhibitors, which include irinotecan and topotecan, are agents that interact with the DNA-topoisomerase I complex and avert resealing of the DNA. The accretion of DNA breaks leads to the inhibition of DNA replication and cell death. On the other hand, topoisomerase II inhibitors like etoposide and teniposide, function by cleaving the DNA-topoisomerase II complex thereby effectively impeding the release of double-strand DNA breaks. Moreover, the recent advances in exploring the therapeutic efficacy, toxicity, and MDR (multidrug resistance) issues of new topoisomerase inhibitors have been reviewed in the present review. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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29. To Break or Not to Break: The Role of TOP2B in Transcription.
- Author
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Cowell, Ian G., Casement, John W., and Austin, Caroline A.
- Subjects
- *
DNA topoisomerase I , *DNA topoisomerase II , *DOUBLE-strand DNA breaks , *DEOXYRIBOZYMES , *DNA topoisomerases , *GENETIC regulation - Abstract
Transcription and its regulation pose challenges related to DNA torsion and supercoiling of the DNA template. RNA polymerase tracking the helical groove of the DNA introduces positive helical torsion and supercoiling upstream and negative torsion and supercoiling behind its direction of travel. This can inhibit transcriptional elongation and other processes essential to transcription. In addition, chromatin remodeling associated with gene activation can generate or be hindered by excess DNA torsional stress in gene regulatory regions. These topological challenges are solved by DNA topoisomerases via a strand-passage reaction which involves transiently breaking and re-joining of one (type I topoisomerases) or both (type II topoisomerases) strands of the phosphodiester backbone. This review will focus on one of the two mammalian type II DNA topoisomerase enzymes, DNA topoisomerase II beta (TOP2B), that have been implicated in correct execution of developmental transcriptional programs and in signal-induced transcription, including transcriptional activation by nuclear hormone ligands. Surprisingly, several lines of evidence indicate that TOP2B-mediated protein-free DNA double-strand breaks are involved in signal-induced transcription. We discuss the possible significance and origins of these DSBs along with a network of protein interaction data supporting a variety of roles for TOP2B in transcriptional regulation. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
30. Narciclasine, a novel topoisomerase I inhibitor, exhibited potent anti-cancer activity against cancer cells.
- Author
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Wang, Meichen, Liang, Leilei, Wang, Rong, Jia, Shutao, Xu, Chang, Wang, Yuting, Luo, Min, Lin, Qiqi, Yang, Min, Zhou, Hongyu, Liu, Dandan, and Qing, Chen
- Subjects
DNA topoisomerase I ,CANCER cells ,DNA topoisomerases ,ANTINEOPLASTIC agents ,DRUG discovery ,CELL cycle - Abstract
DNA topoisomerases are essential nuclear enzymes in correcting topological DNA errors and maintaining DNA integrity. Topoisomerase inhibitors are a significant class of cancer chemotherapeutics with a definite curative effect. Natural products are a rich source of lead compounds for drug discovery, including anti-tumor drugs. In this study, we found that narciclasine (NCS), an amaryllidaceae alkaloid, is a novel inhibitor of topoisomerase I (topo I). Our data demonstrated that NCS inhibited topo I activity and reversed its unwinding effect on p-HOT DNA substrate. However, it had no obvious effect on topo II activity. The molecular mechanism of NCS inhibited topo I showed that NCS did not stabilize topo-DNA covalent complexes in cells, indicating that NCS is not a topo I poison. A blind docking result showed that NCS could bind to topo I, suggesting that NCS might be a topo I suppressor. Additionally, NCS exhibited a potent anti-proliferation effect in various cancer cells. NCS arrested the cell cycle at G
2 /M phase and induced cell apoptosis. Our study reveals the antitumor mechanisms of NCS and provides a good foundation for the development of anti-cancer drugs based on topo I inhibition. [ABSTRACT FROM AUTHOR]- Published
- 2023
- Full Text
- View/download PDF
31. Inhibition of Topoisomerases by Metal Thiosemicarbazone Complexes.
- Author
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Jiang, Xiaohua, Fielding, Lauren A., Davis, Hunter, Carroll, William, Lisic, Edward C., and Deweese, Joseph E.
- Subjects
- *
DNA topoisomerase I , *THIOSEMICARBAZONES , *DEOXYRIBOZYMES , *METAL complexes , *DNA replication , *DRUG target , *ANTINEOPLASTIC agents - Abstract
Topoisomerases, common targets for anti-cancer therapeutics, are crucial enzymes for DNA replication, transcription, and many other aspects of DNA metabolism. The potential anti-cancer effects of thiosemicarbazones (TSC) and metal–TSC complexes have been demonstrated to target several biological processes, including DNA metabolism. Human topoisomerases were discovered among the molecular targets for TSCs, and metal-chelated TSCs specifically displayed significant inhibition of topoisomerase II. The processes by which metal–TSCs or TSCs inhibit topoisomerases are still being studied. In this brief review, we summarize the TSCs and metal–TSCs that inhibit various types of human topoisomerases, and we note some of the key unanswered questions regarding this interesting class of diverse compounds. [ABSTRACT FROM AUTHOR]
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- 2023
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32. Recent advances of benzimidazole as anticancer agents.
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Venugopal, Sneha, Kaur, Balwinder, Verma, Anil, Wadhwa, Pankaj, Magan, Muskan, Hudda, Sharwan, and Kakoty, Violina
- Subjects
- *
ANTINEOPLASTIC agents , *NUCLEIC acids , *TUBULINS , *BENZIMIDAZOLES , *ANDROGEN receptors , *ELECTROSTATIC interaction - Abstract
Cancer is the second leading cause of death globally, with 9.6 million deaths yearly. As a life‐threatening disease, it necessitates the emergence of new therapies. Resistance to current chemotherapies drives scientists to develop new medications that will eventually be accessible. Because heterocycles are so common in biological substances, compounds play a big part in the variety of medications that have been developed. The "Master Key" is the benzimidazole nucleus, which consists of a six‐membered benzene ring fused with a five‐membered imidazole/imidazoline ring, which is an azapyrrole. One of the five‐membered aromatic nitrogen heterocycles identified in American therapies that have been approved by the Food and Drug Administration (FDA). Our results show that benzimidazole's broad therapeutic spectrum is due to its structural isosteres with purine, which improves hydrogen bonding, electrostatic interactions with topoisomerase complexes, intercalation with DNA, and other functions. It also enhances protein and nucleic acid inhibition, tubulin microtubule degeneration, apoptosis, DNA fragmentation, and other functions. Additionally, readers for designing the more recent benzimidazole analogues as prospective cancer treatments. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
33. TOP1 inhibition therapy protects against SARS-CoV-2-induced lethal inflammation
- Author
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Ho, Jessica Sook Yuin, Mok, Bobo Wing-Yee, Campisi, Laura, Jordan, Tristan, Yildiz, Soner, Parameswaran, Sreeja, Wayman, Joseph A, Gaudreault, Natasha N, Meekins, David A, Indran, Sabarish V, Morozov, Igor, Trujillo, Jessie D, Fstkchyan, Yesai S, Rathnasinghe, Raveen, Zhu, Zeyu, Zheng, Simin, Zhao, Nan, White, Kris, Ray-Jones, Helen, Malysheva, Valeriya, Thiecke, Michiel J, Lau, Siu-Ying, Liu, Honglian, Zhang, Anna Junxia, Lee, Andrew Chak-Yiu, Liu, Wen-Chun, Jangra, Sonia, Escalera, Alba, Aydillo, Teresa, Melo, Betsaida Salom, Guccione, Ernesto, Sebra, Robert, Shum, Elaine, Bakker, Jan, Kaufman, David A, Moreira, Andre L, Carossino, Mariano, Balasuriya, Udeni BR, Byun, Minji, Albrecht, Randy A, Schotsaert, Michael, Garcia-Sastre, Adolfo, Chanda, Sumit K, Miraldi, Emily R, Jeyasekharan, Anand D, TenOever, Benjamin R, Spivakov, Mikhail, Weirauch, Matthew T, Heinz, Sven, Chen, Honglin, Benner, Christopher, Richt, Juergen A, and Marazzi, Ivan
- Subjects
Biomedical and Clinical Sciences ,Clinical Sciences ,Biodefense ,Infectious Diseases ,Vaccine Related ,Pneumonia ,Emerging Infectious Diseases ,Lung ,Pneumonia & Influenza ,Prevention ,Development of treatments and therapeutic interventions ,5.1 Pharmaceuticals ,Inflammatory and immune system ,Good Health and Well Being ,Animals ,COVID-19 ,Chlorocebus aethiops ,DNA Topoisomerases ,Type I ,Humans ,Inflammation ,Mesocricetus ,Mice ,Mice ,Transgenic ,SARS-CoV-2 ,THP-1 Cells ,Topoisomerase I Inhibitors ,Topotecan ,Vero Cells ,COVID-19 Drug Treatment ,chromatin ,cytokine storm ,epigenetics ,inducible genes ,inflammation ,topoisomerase ,topotecan ,transcription ,Biological Sciences ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences - Abstract
The ongoing pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is currently affecting millions of lives worldwide. Large retrospective studies indicate that an elevated level of inflammatory cytokines and pro-inflammatory factors are associated with both increased disease severity and mortality. Here, using multidimensional epigenetic, transcriptional, in vitro, and in vivo analyses, we report that topoisomerase 1 (TOP1) inhibition suppresses lethal inflammation induced by SARS-CoV-2. Therapeutic treatment with two doses of topotecan (TPT), an FDA-approved TOP1 inhibitor, suppresses infection-induced inflammation in hamsters. TPT treatment as late as 4 days post-infection reduces morbidity and rescues mortality in a transgenic mouse model. These results support the potential of TOP1 inhibition as an effective host-directed therapy against severe SARS-CoV-2 infection. TPT and its derivatives are inexpensive clinical-grade inhibitors available in most countries. Clinical trials are needed to evaluate the efficacy of repurposing TOP1 inhibitors for severe coronavirus disease 2019 (COVID-19) in humans.
- Published
- 2021
34. Variation of Structure and Cellular Functions of Type IA Topoisomerases across the Tree of Life
- Author
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Kemin Tan and Yuk-Ching Tse-Dinh
- Subjects
topoisomerase ,type IA ,supercoiling ,genome topology ,genomic instability ,genetic diseases ,Cytology ,QH573-671 - Abstract
Topoisomerases regulate the topological state of cellular genomes to prevent impediments to vital cellular processes, including replication and transcription from suboptimal supercoiling of double-stranded DNA, and to untangle topological barriers generated as replication or recombination intermediates. The subfamily of type IA topoisomerases are the only topoisomerases that can alter the interlinking of both DNA and RNA. In this article, we provide a review of the mechanisms by which four highly conserved N-terminal protein domains fold into a toroidal structure, enabling cleavage and religation of a single strand of DNA or RNA. We also explore how these conserved domains can be combined with numerous non-conserved protein sequences located in the C-terminal domains to form a diverse range of type IA topoisomerases in Archaea, Bacteria, and Eukarya. There is at least one type IA topoisomerase present in nearly every free-living organism. The variation in C-terminal domain sequences and interacting partners such as helicases enable type IA topoisomerases to conduct important cellular functions that require the passage of nucleic acids through the break of a single-strand DNA or RNA that is held by the conserved N-terminal toroidal domains. In addition, this review will exam a range of human genetic disorders that have been linked to the malfunction of type IA topoisomerase.
- Published
- 2024
- Full Text
- View/download PDF
35. Cryo-EM structures of African swine fever virus topoisomerase
- Author
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Yan Zhao, Wenhua Kuang, Qiyin An, Jinyue Li, Yong Wang, and Zengqin Deng
- Subjects
African swine fever virus ,cryo-EM ,topoisomerase ,P1192R ,drug target ,Microbiology ,QR1-502 - Abstract
ABSTRACT Type II topoisomerases ubiquitously exist in cellular organisms, where they play an essential role in resolving the topological problems of DNA. The viral type II topoisomerase encoded by the African swine fever virus (ASFV) is critical for viral replication and infection, thus representing an attractive target for antiviral drug development. Here we report two cryo-EM structures of ASFV topoisomerase P1192R in distinct conformations at an overall resolution of 3.16 Å and 3.13 Å, respectively. P1192R assembles as a homodimer with the C-gate formed by the coiled-coil domain adopting a closed or open conformation before reaction, providing the first visual evidence for the dynamic motions of the C-gate of type II topoisomerase. Comparative structural comparisons of eukaryotic homologs and P1192R reveal the unique structural features of P1192R, including the active site configuration, a flexible loop in the TOPRIM domain, an additionally inserted α-helix in the tower domain, and a pin-like structure in the C-terminal coiled-coil domain, which are important for enzyme activity and protein folding. These findings provide important insights into the structure and function of viral topoisomerases and may guide the efficient development of anti-ASFV drugs. Moreover, our study also offers structural evidence to support the scenario of the viral origin of eukaryotic type IIA topoisomerases. IMPORTANCE African swine fever virus (ASFV) is a highly contagious virus that causes lethal hemorrhagic diseases known as African swine fever (ASF) with a case fatality rate of 100%. There is an urgent need to develop anti-ASFV drugs. We determine the first high-resolution structures of viral topoisomerase ASFV P1192R in both the closed and open C-gate forms. P1192R shows a similar overall architecture with eukaryotic and prokaryotic type II topoisomerases, which have been successful targets of many antimicrobials and anticancer drugs, with the most similarity to yeast topo II. P1192R also exhibits differences in the details of active site configuration, which are important to enzyme activity. These two structures offer useful structural information for antiviral drug design and provide structural evidence to support that eukaryotic type IIA topoisomerase likely originated from horizontal gene transfer from the virus.
- Published
- 2023
- Full Text
- View/download PDF
36. Dynamics of chromosome organization in a minimal bacterial cell
- Author
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Benjamin R. Gilbert, Zane R. Thornburg, Troy A. Brier, Jan A. Stevens, Fabian Grünewald, John E. Stone, Siewert J. Marrink, and Zaida Luthey-Schulten
- Subjects
whole-cell modeling ,chromosome replication ,chromosome segregation ,brownian dynamics ,smc proteins ,topoisomerase ,Biology (General) ,QH301-705.5 - Abstract
Computational models of cells cannot be considered complete unless they include the most fundamental process of life, the replication and inheritance of genetic material. By creating a computational framework to model systems of replicating bacterial chromosomes as polymers at 10 bp resolution with Brownian dynamics, we investigate changes in chromosome organization during replication and extend the applicability of an existing whole-cell model (WCM) for a genetically minimal bacterium, JCVI-syn3A, to the entire cell-cycle. To achieve cell-scale chromosome structures that are realistic, we model the chromosome as a self-avoiding homopolymer with bending and torsional stiffnesses that capture the essential mechanical properties of dsDNA in Syn3A. In addition, the conformations of the circular DNA must avoid overlapping with ribosomes identitied in cryo-electron tomograms. While Syn3A lacks the complex regulatory systems known to orchestrate chromosome segregation in other bacteria, its minimized genome retains essential loop-extruding structural maintenance of chromosomes (SMC) protein complexes (SMC-scpAB) and topoisomerases. Through implementing the effects of these proteins in our simulations of replicating chromosomes, we find that they alone are sufficient for simultaneous chromosome segregation across all generations within nested theta structures. This supports previous studies suggesting loop-extrusion serves as a near-universal mechanism for chromosome organization within bacterial and eukaryotic cells. Furthermore, we analyze ribosome diffusion under the influence of the chromosome and calculate in silico chromosome contact maps that capture inter-daughter interactions. Finally, we present a methodology to map the polymer model of the chromosome to a Martini coarse-grained representation to prepare molecular dynamics models of entire Syn3A cells, which serves as an ultimate means of validation for cell states predicted by the WCM.
- Published
- 2023
- Full Text
- View/download PDF
37. Naturally mutagenic sequence diversity in a human type II topoisomerase.
- Author
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Bandak, Afif F., Blower, Tim R., Nitiss, Karin C., Gupta, Raveena, Lau, Albert Y., Guha, Ria, Nitiss, John L., and Berger, James M.
- Subjects
- *
DNA topoisomerase II , *GENETIC testing , *CELL transformation , *MUTAGENS , *MOLECULAR dynamics - Abstract
Type II topoisomerases transiently cleave duplex DNA as part of a strand passage mech- anism that helps control chromosomal organization and superstructure. Aberrant DNA cleavage can result in genomic instability, and how topoisomerase activity is controlled to prevent unwanted breaks is poorly understood. Using a genetic screen, we identified mutations in the beta isoform of human topoisomerase II (hTOP2β) that render the enzyme hypersensitive to the chemotherapeutic agent etoposide. Several of these variants were unexpectedly found to display hypercleavage behavior in vitro and to be capable of inducing cell lethality in a DNA repair–deficient background; surprisingly, a subset of these mutations were also observed in TOP2B sequences from cancer genome databases. Using molecular dynamics simulations and computational network analyses, we found that many of the mutations obtained from the screen map to interfacial points between structurally coupled elements, and that dynamical modeling could be used to identify other damage-inducing TOP2B alleles present in cancer genome databases. This work establishes that there is an innate link between DNA cleavage predisposition and sensitivity to topoisomerase II poisons, and that certain sequence variants of human type II topoi- somerases found in cancer cells can act as DNA-damaging agents. Our findings underscore the potential for hTOP2β to function as a clastogen capable of generating DNA damage that may promote or support cellular transformation. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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38. Design and in silico Prediction of New Methylenedioxyphenyl Derivatives as DNA Topoisomerase II Inhibitors.
- Author
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Rathore, Swati, Reddemma, Maadhu, Rai, Neha, Tiwari, Richa Tripathy, Begum, Shaheen, Patil, Shailendra, and Patil, Asmita Gajbhiye
- Subjects
- *
DNA topoisomerase I , *DNA topoisomerase II , *DNA topoisomerase inhibitors , *MOLECULAR docking , *CAUSES of death - Abstract
Background: Cancer is the world's leading cause of death; more than ten million people die from cancer each year. Rate of morbidity and mortality is increasing day by day. As a result, utilizing chemoinformatics techniques, a novel series of Methylenedioxyphenyl linked with 3,4,5-trimethoxyphenyl/2,5-dimethoxyphenyl and pyrimidine has been designed, docked and in-silico predictions of pharmacokinetic, and toxicological parameters. Materials and Methods: A novel class of Methylenedioxyphenyl derivatives was docked by using AutoDock Vina software to reveal the interaction of these derivatives with the active site topoisomerase-II with PBD ID 5GWK. In addition to the above, the ADME studies were performed using Swiss ADME software and the toxicity parameters using the pkCSM online tool. Results: Many derivatives were found to be more active than the standard drug etoposide in docking studies. Binding interaction pattern of selected compounds were studied with the active sites of DNA topoisomerase-II (PBD ID 5GWK) by docking simulation. The results of the screening revealed that compounds B9, B12, B13, B15, B16, B17, B18, B22, and B23 were more active candidates of the series. Conclusion: Molecular docking and ADME/Tox properties of new methylenedioxyphenyl derivatives have been described. As a result, nine compounds from the intended series showed improved docking scores with suitable ADME/Tox properties and satisfactory topo II inhibiting activity, thus these compounds may be the effective inhibitors of topoisomerase IIa enzyme. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
39. Mechanical determinants of chromatin topology and gene expression
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Rajiv Kumar Jha, David Levens, and Fedor Kouzine
- Subjects
Transcription ,chromatin ,psoralen ,DNA supercoiling ,3D genome ,topoisomerase ,Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
The compaction of linear DNA into micrometer-sized nuclear boundaries involves the establishment of specific three-dimensional (3D) DNA structures complexed with histone proteins that form chromatin. The resulting structures modulate essential nuclear processes such as transcription, replication, and repair to facilitate or impede their multi-step progression and these contribute to dynamic modification of the 3D-genome organization. It is generally accepted that protein–protein and protein–DNA interactions form the basis of 3D-genome organization. However, the constant generation of mechanical forces, torques, and other stresses produced by various proteins translocating along DNA could be playing a larger role in genome organization than currently appreciated. Clearly, a thorough understanding of the mechanical determinants imposed by DNA transactions on the 3D organization of the genome is required. We provide here an overview of our current knowledge and highlight the importance of DNA and chromatin mechanics in gene expression.
- Published
- 2022
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40. From Antarctica to cancer research: a novel human DNA topoisomerase 1B inhibitor from Antarctic sponge Dendrilla antarctica
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Alessio Ottaviani, Joshua Welsch, Keli Agama, Yves Pommier, Alessandro Desideri, Bill J. Baker, and Paola Fiorani
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Natural product ,topoisomerase ,cancer ,drug development ,Therapeutics. Pharmacology ,RM1-950 - Abstract
Nature has been always a great source of possible lead compounds to develop new drugs against several diseases. Here we report the identification of a natural compound, membranoid G, derived from the Antarctic sponge Dendrilla antarctica displaying an in vitro inhibitory activity against human DNA topoisomerase 1B. The experiments indicate that membranoid G, when pre-incubated with the enzyme, strongly and irreversibly inhibits the relaxation of supercoiled DNA. This compound completely inhibits the cleavage step of the enzyme catalytic mechanism by preventing protein binding to the DNA. Membranoid G displays also a cytotoxic effect on tumour cell lines, suggesting its use as a possible lead compound to develop new anticancer drugs.
- Published
- 2022
- Full Text
- View/download PDF
41. Investigation of the chromatin remodelling enzyme Uls1 and its interactions with Topoisomerase 2 in S. cerevisiae
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Swanston, Amy and Ferreira, Helder
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572 ,Molecular biology ,Cell biology ,Top2 ,Uls1 ,Topoisomerase ,Chromatin remodelling ,Chromatin remodeller ,Chromatin ,DNA ,SUMO ,Yeast ,Protein modification ,Acriflavine ,Top2 poison ,Drug screen ,Genetic screen ,Chromatin immunoprecipitation ,ChIP-seq ,Bioinformatics ,QP616.D56S8 ,DNA topoisomerase II ,Adenosine triphosphatase ,Saccharomyces cerevisiae - Abstract
Acriflavine (ACF) is a Topoisomerase 2 (Top2) poison, a class of drugs which stall Top2 during its reaction cycle causing the formation of persistent DNA breaks to which Top2 remains covalently bound. Deletion of ULS1 causes sensitivity to ACF, with cells showing activation of the Rad53 DNA damage checkpoint. Uls1 is a chromatin remodelling enzyme also implicated in the regulation of levels of SUMO conjugated proteins. We show that Uls1 has both a genetic and physical interaction with Top2, with uls1Δ sensitivity to ACF being linked to Top2 activity. Analysis of Uls1 and Top2 localisation genome wide via ChIP-seq reveals areas where the two proteins co-localise, with Top2 enrichment on chromatin being altered upon deletion of ULS1. At these areas, the presence of Uls1 prevents accumulation of Top2 upon addition of ACF. Our data suggests that Uls1 is required for regulation of stalled Top2. Top2 poisons are used therapeutically as anti-cancer drugs, however these drugs have been implicated in the formation of secondary cancers due to chromosomal translocations arising during the repair of Top2 generated double strand breaks (DSB). The use of dual targeted therapies where a Top2 poison is paired with an inhibitor of another pathway that increases sensitivity to the Top2 poison allows a lower dose to be used, therefore reducing harmful side effects. Our work looked to identify Top2 poison sensitive pathways in S. cerevisiae, where non-essential and essential gene mutants were assayed for sensitivity to ACF. This allowed a comprehensive analysis of 83% of the genes in S. cerevisiae, identifying novel genes within the areas of DNA repair, DNA replication, transcription, chromatin structure, protein modification/degradation, cell division/cell cycle and cellular organisation/cytoskeleton as being important in the response to this bulky adduct.
- Published
- 2019
- Full Text
- View/download PDF
42. A nuclear pore complex‐associated regulation of SUMOylation in meiosis.
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Yang, Hui‐Ju, Asakawa, Haruhiko, Li, Fu‐An, Haraguchi, Tokuko, Shih, Hsiu‐Ming, and Hiraoka, Yasushi
- Subjects
- *
DNA topoisomerase I , *MEIOSIS , *DNA topoisomerase II , *NUCLEAR proteins , *SCHIZOSACCHAROMYCES pombe , *AQUAPORINS - Abstract
The nuclear pore complex (NPC) provides a permeable barrier between the nucleoplasm and cytoplasm. In a subset of NPC constituents that regulate meiosis in the fission yeast Schizosaccharomyces pombe, we found that nucleoporin Nup132 (homolog of human Nup133) deficiency resulted in transient leakage of nuclear proteins during meiosis I, as observed in the nup132 gene‐deleted mutant. The nuclear protein leakage accompanied the liberation of the small ubiquitin‐like modifier (SUMO)‐specific ubiquitin‐like protease 1 (Ulp1) from the NPC. Ulp1 retention at the nuclear pore prevented nuclear protein leakage and restored normal meiosis in a mutant lacking Nup132. Furthermore, using mass spectrometry analysis, we identified DNA topoisomerase 2 (Top2) and RCC1‐related protein (Pim1) as the target proteins for SUMOylation. SUMOylation levels of Top2 and Pim1 were altered in meiotic cells lacking Nup132. HyperSUMOylated Top2 increased the binding affinity at the centromeres of nup132 gene‐deleted meiotic cells. The Top2‐12KR sumoylation mutant was less localized to the centromeric regions. Our results suggest that SUMOylation of chromatin‐binding proteins is regulated by the NPC‐bound SUMO‐specific protease and is important for the progression of meiosis. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
43. MYC: a complex problem.
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Das, Subhendu K., Lewis, Brian A., and Levens, David
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- *
POST-translational modification , *REGULATOR genes , *PROMOTERS (Genetics) , *JOINING processes , *STRAINS & stresses (Mechanics) , *ONCOGENES , *DNA replication , *GENE regulatory networks - Abstract
The MYC protooncogene functions as a universal amplifier of transcription through interaction with numerous factors and complexes that regulate almost every cellular process. However, a comprehensive model that explains MYC's actions and the interplay governing the complicated dynamics of components of the transcription and replication machinery is still lacking. Here, we review the potency of MYC as an oncogenic driver and how it regulates the broad spectrum of complexes (effectors and regulators). We propose a 'hand-over model' for differential partitioning and trafficking of unstructured MYC via a loose interaction network between various gene-regulatory complexes and factors. Additionally, the article discusses how unstructured-MYC energetically favors efficient modulation of the energy landscape of the transcription cycle. As an oncogenic driver, MYC amplifies global transcription by driving pause release and the early transcription, especially of highly expressed genes. It stimulates replication by facilitating assembly of replication complexes at origins and by associating with replication forks. It facilitates these processes by joining with a wide variety of gene regulatory complexes. Beyond helping to recruit the transcription machinery to promoters, MYC directly up- or down-modulates the catalytic activities of its interaction partners. For example, through assembly with topoisomerases 1 and 2 in the topoisome, MYC dramatically augments the cell's capacity to confront the topological and conformational challenges of DNA and chromatin under high-output mechanical stress. To orchestrate its activities, MYC levels must be strictly tuned by a series of interdependent enzymes that are responsible for its post-translational modifications, stabilization, destabilization, and trafficking. To coordinate the flux of MYC with its associated complexes, its unstructured regions associate with effectors and complexes at transcription start sites (TSSs) at different stages throughout the transcription cycle. To increase overall transcription output, MYC alters the residence times of components of the transcription machinery and as different complexes are sequentially ferried in and out of promoter regions. The conformational plasticity of MYC's effector regions allow it to adapt to a variety of partners without a prior energetic expense of unfolding or remodeling. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
44. Developments in Non-Intercalating Bacterial Topoisomerase Inhibitors: Allosteric and ATPase Inhibitors of DNA Gyrase and Topoisomerase IV.
- Author
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Grossman, Scott, Fishwick, Colin W. G., and McPhillie, Martin J.
- Subjects
- *
DNA topoisomerase I , *DNA topoisomerase inhibitors , *ADENOSINE triphosphatase , *ANTI-infective agents , *DRUG resistance in bacteria , *SINGLE nucleotide polymorphisms , *ANTIBIOTICS - Abstract
Increases in antibiotic usage and antimicrobial resistance occurrence have caused a dramatic reduction in the effectiveness of many frontline antimicrobial treatments. Topoisomerase inhibitors including fluoroquinolones are broad-spectrum antibiotics used to treat a range of infections, which stabilise a topoisomerase-DNA cleavage complex via intercalation of the bound DNA. However, these are subject to bacterial resistance, predominantly in the form of single-nucleotide polymorphisms in the active site. Significant research has been undertaken searching for novel bioactive molecules capable of inhibiting bacterial topoisomerases at sites distal to the fluoroquinolone binding site. Notably, researchers have undertaken searches for anti-infective agents that can inhibit topoisomerases through alternate mechanisms. This review summarises work looking at the inhibition of topoisomerases predominantly through non-intercalating agents, including those acting at a novel allosteric site, ATPase domain inhibitors, and those offering unique binding modes and mechanisms of action. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
45. Role of the Water–Metal Ion Bridge in Quinolone Interactions with Escherichia coli Gyrase.
- Author
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Carter, Hannah E., Wildman, Baylee, Schwanz, Heidi A., Kerns, Robert J., and Aldred, Katie J.
- Subjects
- *
ESCHERICHIA coli , *BACILLUS anthracis , *GRAM-negative bacteria , *IONS , *DRUG design - Abstract
Fluoroquinolones are an important class of antibacterials, and rising levels of resistance threaten their clinical efficacy. Gaining a more full understanding of their mechanism of action against their target enzymes—the bacterial type II topoisomerases gyrase and topoisomerase IV—may allow us to rationally design quinolone-based drugs that overcome resistance. As a step toward this goal, we investigated whether the water–metal ion bridge that has been found to mediate the major point of interaction between Escherichia coli topoisomerase IV and Bacillus anthracis topoisomerase IV and gyrase, as well as Mycobacterium tuberculosis gyrase, exists in E. coli gyrase. This is the first investigation of the water–metal ion bridge and its function in a Gram-negative gyrase. Evidence suggests that the water–metal ion bridge does exist in quinolone interactions with this enzyme and, unlike the Gram-positive B. anthracis gyrase, does use both conserved residues (serine and acidic) as bridge anchors. Furthermore, this interaction appears to play a positioning role. These findings raise the possibility that the water–metal ion bridge is a universal point of interaction between quinolones and type II topoisomerases and that it functions primarily as a binding contact in Gram-positive species and primarily as a positioning interaction in Gram-negative species. Future studies will explore this possibility. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
46. Dynamic Processing of Displacement Loops during Recombinational DNA Repair
- Author
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Piazza, Aurèle, Shah, Shanaya Shital, Wright, William Douglass, Gore, Steven K, Koszul, Romain, and Heyer, Wolf-Dietrich
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Biotechnology ,Genetics ,Underpinning research ,1.1 Normal biological development and functioning ,DEAD-box RNA Helicases ,DNA Damage ,DNA Helicases ,DNA Topoisomerases ,DNA ,Fungal ,Kinetics ,Nucleic Acid Conformation ,RecQ Helicases ,Recombinational DNA Repair ,Saccharomyces cerevisiae ,Saccharomyces cerevisiae Proteins ,Structure-Activity Relationship ,D-loop ,Mph1 ,Rad54 ,Rdh54 ,Sgs1 ,Srs2 ,helicase ,heteroduplex DNA ,homologous recombination ,topoisomerase ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences ,Health sciences - Abstract
Displacement loops (D-loops) are pivotal intermediates of homologous recombination (HR), a universal DNA double strand break (DSB) repair pathway. We developed a versatile assay for the physical detection of D-loops in vivo, which enabled studying the kinetics of their formation and defining the activities controlling their metabolism. Nascent D-loops are detected within 2 h of DSB formation and extended in a delayed fashion in a genetic system designed to preclude downstream repair steps. The majority of nascent D-loops are disrupted by two pathways: one supported by the Srs2 helicase and the other by the Mph1 helicase and the Sgs1-Top3-Rmi1 helicase-topoisomerase complex. Both pathways operate without significant overlap and are delineated by the Rad54 paralog Rdh54 in an ATPase-independent fashion. This study uncovers a layer of quality control of HR relying on nascent D-loop dynamics.
- Published
- 2019
47. Resistance rate and mechanism of ureaplasma urealyticum to fluoroquinolones in infertile patients
- Author
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YUAN Yaling and XIA Yun
- Subjects
ureaplasma urealyticum ,fluoroquinolones ,topoisomerase ,resistance ,Medicine (General) ,R5-920 - Abstract
Objective To investigate the resistance of ureaplasma urealyticum (Uu) isolated from infertile patients to fluoroquinolones, and explore the correlation between quinolone resistance determining regions (QRDR) mutations in topoisomerase gene and antimicrobial resistance. Methods Clinical strains of ureaplasma isolated and cultured from the genitourinary tract of infertile patients who were admitted between December 2019 and January 2022 were collected, and validated by 16SrRNA sequencing. Two conserved genes, UU295 and UUR10_0588, were subsequently adopted to classify the strains into ureaplasma parvum (UPA) and ureaplasma urealyticum (UUR). In addition, the minimum inhibitory concentrations of ciprofloxacin, levofloxacin, sparfloxacin and moxifloxacin were determined by broth dilution method. The resistant strains were subsequently screened according to CLSI M43-A (2011 edition) standard, topoisomerase gene QRDR was amplified by PCR, and the sequencing results were analyzed using DNASTAR Lasergene. Finally, the homology models of parC S83L and gyrB R523E mutations were performed by Swiss-model homology modeling server. Results A total of 184 clinical strains were isolated, including 173 UPA (94.0%), 8 UUR (4.4%), and 3 of co-existing (1.6%). The resistance rates of the 3 groups to levofloxacin and moxifloxacin were UPA (80.4%, 2.2%), UUR (50%, 0), and UPA+UUR (66.7%, 33.3%), respectively. The incidence of QRDR amino acid variants was parC S83L (85.3%), gyrB R523E (11.4%), parC L127P (7.1%), gyrA L176F (2.2%), gyrA S174Q (1.6%), and parE L408I (1.1%), among which parE L408I is a newly discovered mutation. Homology modeling revealed that the parC S83L mutation interferes with the normal binding of quinolones by forming spatial hindrance, leading to fluoroquinolone resistance. Conclusion The resistance rate of Uu strains to fluoroquinolones in infertile patients is rather high, which is closely related to the amino acid variation caused by the mutation of topoisomerase gene parC S83L base.
- Published
- 2022
- Full Text
- View/download PDF
48. Target-based anticancer indole derivatives and insight into structure‒activity relationship: A mechanistic review update (2018–2021)
- Author
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Ashima Dhiman, Rupam Sharma, and Rajesh K. Singh
- Subjects
Indole ,Synthesis ,Anticancer ,Structure‒activity relationship ,Topoisomerase ,Apoptosis ,Therapeutics. Pharmacology ,RM1-950 - Abstract
Cancer, which is the uncontrolled growth of cells, is the second leading cause of death after heart disease. Targeting drugs, especially to specific genes and proteins involved in growth and survival of cancer cells, is the prime need of research world-wide. Indole moiety, which is a combination of aromatic-heterocyclic compounds, is a constructive scaffold for the development of novel leads. Owing to its bioavailability, high unique chemical properties and significant pharmacological behaviours, indole is considered as the most inquisitive scaffold for anticancer drug research. This is illustrated by the fact that the U.S. Food and Drug Administration (FDA) has recently approved several indole-based anticancer agents such as panobinostat, alectinib, sunitinib, osimertinib, anlotinib and nintedanib for clinical use. Furthermore, hundreds of studies on the synthesis and activity of the indole ring have been published in the last three years. Taking into account the facts stated above, we have presented the most recent advances in medicinal chemistry of indole derivatives, encompassing hot articles published between 2018 and 2021 in anticancer drug research. The recent advances made towards the synthesis of promising indole-based anticancer compounds that may act via various targets such as topoisomerase, tubulin, apoptosis, aromatase, kinases, etc., have been discussed. This review also summarizes some of the recent efficient green chemical synthesis for indole rings using various catalysts for the period during 2018–2021. The review also covers the synthesis, structure‒activity relationship, and mechanism by which these leads have demonstrated improved and promising anticancer activity. Indole molecules under clinical and preclinical stages are classified into groups based on their cancer targets and presented in tabular form, along with their mechanism of action. The goal of this review article is to point the way for medicinal chemists to design and develop effective indole-based anticancer agents.
- Published
- 2022
- Full Text
- View/download PDF
49. A sumoylation program is essential for maintaining the mitotic fidelity in proliferating mantle cell lymphoma cells
- Author
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Walter Hanel, Pushpa Lata, Youssef Youssef, Ha Tran, Liudmyla Tsyba, Lalit Sehgal, Bradley W. Blaser, Dennis Huszar, JoBeth Helmig-Mason, Liwen Zhang, Morgan S. Schrock, Matthew K. Summers, Wing Keung Chan, Alexander Prouty, Bethany L. Mundy-Bosse, Selina Chen-Kiang, Alexey V. Danilov, Kami Maddocks, Robert A. Baiocchi, and Lapo Alinari
- Subjects
UBA2 ,SAE1 ,SAE2 ,UBC9 ,UBE2I ,Topoisomerase ,Diseases of the blood and blood-forming organs ,RC633-647.5 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Abstract Background Mantle cell lymphoma (MCL) is a rare, highly heterogeneous type of B-cell non-Hodgkin’s lymphoma. The sumoylation pathway is known to be upregulated in many cancers including lymphoid malignancies. However, little is known about its oncogenic role in MCL. Methods Levels of sumoylation enzymes and sumoylated proteins were quantified in MCL cell lines and primary MCL patient samples by scRNA sequencing and immunoblotting. The sumoylation enzyme SAE2 was genetically and pharmacologically targeted with shRNA and TAK-981 (subasumstat). The effects of SAE2 inhibition on MCL proliferation and cell cycle were evaluated using confocal microscopy, live-cell microscopy, and flow cytometry. Immunoprecipitation and orbitrap mass spectrometry were used to identify proteins targeted by sumoylation in MCL cells. Results MCL cells have significant upregulation of the sumoylation pathway at the level of the enzymes SAE1 and SAE2 which correlated with poor prognosis and induction of mitosis associated genes. Selective inhibition of SAE2 with TAK-981 results in significant MCL cell death in vitro and in vivo with mitotic dysregulation being an important mechanism of action. We uncovered a sumoylation program in mitotic MCL cells comprised of multiple pathways which could be directly targeted with TAK-981. Centromeric localization of topoisomerase 2A, a gene highly upregulated in SAE1 and SAE2 overexpressing MCL cells, was lost with TAK-981 treatment likely contributing to the mitotic dysregulation seen in MCL cells. Conclusions This study not only validates SAE2 as a therapeutic target in MCL but also opens the door to further mechanistic work to uncover how to best use desumoylation therapy to treat MCL and other lymphoid malignancies.
- Published
- 2022
- Full Text
- View/download PDF
50. Clinical pharmacokinetics and toxicity of irinotecan
- Author
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Attina, G, Mastrangelo, S, and Ruggiero, A
- Published
- 2022
- Full Text
- View/download PDF
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