function of topoisomerase ii

Thus, topoisomerase II introduces a transient double strand break in DNA. Control of topoisomerase II activity and chemotherapeutic ... Cellular distribution of mammalian DNA topoisomerase II is ... Interacts with ERCC6 (PubMed: 26030138 ). Name Topoisomerase II Inhibitors Accession Number DBCAT000549 Description. What is the Difference Between Topoisomerase I and II ... (PDF) Structure and function of type II DNA topoisomerases Mammalian cells express two genetically distinct isoforms of DNA topoisomerase II, designated topoisomerase IIalphaand topoisomerase IIbeta. Differential regulation of NF-κB activation and function ... Functional Expression of a Drosophila Gene in Yeast ... Its procaryotic equivalent, DNA gyrase, is the target of the quinolones, highly successful antibacterial compounds. In cells, type II topoisomerases are particularly useful for their ability to disentangle newly-replicated sister chromosomes. Full size table Daunorubicin and doxorubicin also undergo redox cycling and generate oxygen free radicals, which can have a variety of effects, including damage to cell membranes but also provides a third . Faithful chromosome segregation depends on the precise timing of chromatid separation, which is enforced by checkpoint signals generated at kinetochores. that DNA Topoisomerase II (Topo II) is also required to modulate the activity of the Su(Hw) insulator. DNA Topoisomerases: Structure, Function, and Mechanism ... Topoisomerase II poisons generate DNA damage, in addition to inhibition of enzyme activity, and would be expected to delay cell cycle progression by means of DNA damage checkpoints. ab72334 at 0.1µg/ml for WB. It does not need ATP hydrolyzing to catalyze the topological rearrangement of DNA. Difference Between Topoisomerase I and II | Compare the ... We find that the yeast CTD is required for recruitment of the tension checkpoint kinase Ipl1/Aurora B to inner centromeres in metaphase but is not required in interphase. Clarke, D J, and Y Azuma. Topoisomerase II beta mRNA was expressed in haemopoietic, epithelial and fibroblast cell lines, although to different extents, with U937 cells (promonocytic leukaemia) showing a particularly high . Helicase vs Topoisomerase (DNA gyrase) - Biology Brain Here, we provide evidence that the C-terminal domain (CTD) of DNA topoisomerase IIα (Topo II) provides a novel function at inner centromeres of kinetochores in mitosis. Binding Cleavage Religation & Release 18. These drugs act in an insidious fashion and kill cells by increasing levels of covalent topoisomerase II-cleaved DNA complexes that are normally fleeting . Medical Definition of Topoisomerase - RxList Figure 01: Topoisomerase I and II The Ecoli topoisomerase I is a holoenzyme with three Zn (II) atoms in the tetracysteine motifs near its carboxy terminus. Both representations show only the DNA-binding gate and not the ATPase segment. To this end, four broad-based projects are currently underway. We have isolated cDNA encoding DNA topoisomerase II and have localized the gene on Beyond its physiological functions, topoisomerase II is the target for some of the most active and widely prescribed anticancer drugs currently utilized for the treatment of human cancers. Circular DNA is covalently closed and does not have any interruption in between nucleotides. Dong and Berger have described a structure of the breakage reunion domain of yeast topoisomerase II (Top2) bound to DNA 36.A key feature of the structure is the large bend induced in the DNA. Both isoforms can complement the single form of the enzyme in yeast , but in mammals only Top2α can provide the type II topoisomerase functions required for DNA propagation, such as chromosome condensation and segregation (32, 53). Topoisomerase IIα is a multifunctional enzyme that catalyzes the relaxation of supercoiled DNA, decatenation of interlinked DNA and unknotting of intramolecularly linked DNA by passing a DNA helix through a transient double-strand break in a second helix (1,2).Consistent with its role in chromosome segregation, topoisomerase II was identified as ScI (sc affold-1), a major . All type-I topoisomerases cannot carry out the full range of activities. Interacts with MCM3AP isoform GANP (PubMed: 23652018 ). Here, we provide evidence that the C-terminal domain (CTD) of DNA topoisomerase IIα (Topo II) provides a novel function at inner centromeres of kinetochores in mitosis. A noncatalytic function of the topoisomerase II CTD in Aurora B recruitment to inner centromeres during mitosis Heather Edgerton. 28, Ienue of October 5, pp. Here we report a new function of the mitochondrial topoisomerase II (TbTOP2mt). Chung et al. Therapeutic use of irinotecan. Three forms of DNA is most prevalent in nature: circular, linear and supercoiled.". We are interested in analyzing the functional roles of DNA topoisomerase II during the growth and development of Drosophila melanogaster. We have recently shown that mouse topoisomerase IIalpha can substitute for the yeast topoisomerase II enzyme and complement yeast top2 mutations. In mammalian cells, it is the target of the anthracyclines and epiphyllotoxins, widely used anticancer agents. Topoisomerase: A class of enzymes that alter the supercoiling of double-stranded DNA. In the 1970s, James C. Wang was the first to discover a topoisomerase when he identified E. coli topoisomerase I. Topo EC-codes are as follows: ATP-independent (type I), EC 5.6.2.1; ATP-dependent (Type II): EC 5.6.2.2. Type II topoisomerases increase or decrease the linking number of a DNA loop by 2 units, and it promotes chromosome disentanglement. Topoisomerase type I cuts one strand whereas topoisomerase type II cuts both strands of the DNA to relax the coil and extend the DNA molecule. )The topoisomerases act by transiently cutting one or both strands of the DNA. Topoisomerase II is a universally essential enzyme ( Watt and Hickson, 1994 ). In mammals, there are 2 isoforms of DNA Top II, termed Top IIα and Top IIβ. Topoisomerase IIalpha stably interacted with RNA harboring a 3'-hydroxyl group but not with RNA possessing a 3'-phosphate group. 25 between topo I and topo II enzymes because it allows us to estimate topo II enzyme activity independent of topo I activity in vitro 3.9 Differences between DNA Topoisomerase II and Topo II Topo II has two isozymes, topo II and topo II with different molecular masses and gene loci (Lang et al., 1998; Drake et al ., 1987) (Table 1, page 13). Type II topoisomerase can break DNA in a region of chromosome 11q23 that contains the ataxia telangiectasia gene (ATM). Type IB Topoisomerase Functions via a Controlled Rotation Mechanism 17. The regulation of DNA supercoiling is essential to DNA transcription and replication, when the DNA helix must unwind to permit the proper function of the enzymatic machinery involved in these processes. In the presence of a nonhydrolyzable ATP analog, the enzyme is known to promote a single turnover of DNA transport. LC-MS/MS analysis identifies TCA cycle intermediates as stimulators of topo II activity. 11 (October 17, 2017). Topoisomerase IIα is a multifunctional enzyme that catalyzes the relaxation of supercoiled DNA, decatenation of interlinked DNA and unknotting of intramolecularly linked DNA by passing a DNA helix through a transient double-strand break in a second helix (1,2).Consistent with its role in chromosome segregation, topoisomerase II was identified as ScI (sc affold-1), a major . . They identified 2 classes of sequence, which represent the TOP2 isoenzymes TOP2A and TOP2B. [Source 12)] Topoisomerase 2. The use of temperature-sensitive mutants in topoisomerase II have demonstrated roles in the relaxation of tortional stress, reduction of recombination rates, and in the separation of sister chromatids after replication. There are two types or families of this enzyme; type I . Structural Features. The goal of my laboratory is to define the function and biology of eukaryotic topoisomerase II. Immunoprecipitation - Anti-Topoisomerase II beta/TOP2B antibody (ab72334) Immunoprecipitation/ Western Blot of Topoisomerase II beta/TOP2B. DNA topoisomerase II is a homodimeric molecular machine that couples ATP usage to the transport of one DNA segment through a transient break in another segment. topoisomerase II is actively exported from the nuclease and is mediated by a CRM1-dependent pathway; Topo-II alpha is a useful marker for diagnosing liposarcoma. NX_Q02880 - TOP2B - DNA topoisomerase 2-beta - Function. The break with topoisomerase II has a four base stagger. Topoisomerase II. Function. DNA topoisomerase II (topo II) plays a crucial role in controlling the conformation of both DNA and whole chromosomes. Type II DNA topoisomerases (Tops) are ATP-dependent enzymes that catalyze topological transformations of genomic DNA by the transport of one DNA double helix through another. This activity is essential for several cellular events such as DNA replication, transcription, chromosome condensation and segregation. 268, No. Spacefill of topoisomerase II . Lane 1: ab72334 at 3µg/mg whole cell lysate. Selected Publications. In addition, these enzymes fine-tune the steady-state level of DNA supercoiling both to facilitate protein interactions with the DNA and to prevent excessive supercoiling that is . Compounds that inhibit the activity of DNA TOPOISOMERASE II. Detection: DAB. Mammals have two isoforms of Topoisomerase II with similar enzymatic properties in vitro (7, 8). This . It is 97 kDa in weight. Therapeutic use of topotecan. In vertebrate cells, topoisomerase II was shown to be . Search for other works by this author on: . Kaur, K, H Park, N Pandey, Y Azuma, and R N De Guzman. Mondal and Parvin (2001) demonstrated that DNA topoisomerase II-alpha is associated with the pol II holoenzyme and is a required component of chromatin-dependent coactivation. Relating Structure to Function Through the Dominant Slow Modes of Motion of DNA Topoisomerase II R. L. JERNIGAN,1 M. C. DEMIREL,1,2,3 I. BAHAR1,2,3 1 Molecular Structure Section, Laboratory of Experimental and Computational Biology, Division of Basic Sciences, National Cancer Institute, National Institutes of Health, Although traditionally thought to reattach minicircle progeny to the network, here we show that it also mends holes in the network created by minicircle release. INTRODUCTION. Finally, the cut ends are relegated again. We find that the yeast CTD is required for recruitment of the tension . This . Topoisomerase alpha is showns as a monomer and topoisomerase beta is shown as a dimer. The goal of this work is to analyze both expression and function of topoisomerases during the meiotic stages of mammalian spermatogenesis. Circular DNA is found in . prodrug that is concerted metabolically to active metabolite S…. The topoisomerase can change the topology of DNA by changing the linking number. topoisomerase II α (Topo II) catalyzes this resolving activity and is thus essential for mitosis in all eukaryotes (Nitiss, 2009a). Similarities Between Topoisomerase I and II. Topoisomerase II is a homodimer, which functions by clamping onto DNA, positioning its catalytic residues next to two Mg ions and the DNA backbone. International Journal of Molecular Sciences 18, no. Lee et al. Topo II has been shown to be important for critical cellular functions such as transcription, replication, recombination and genome stability in addition to its function in the organization of chromatin architecture [26,27,28]. It is less clear, however, whether Top2 performs the same function uniformly across the whole genome, and whether all its functions rely on decatenation. Relating Structure to Function Through the Dominant Slow Modes of Motion of DNA Topoisomerase II R. L. JERNIGAN,1 M. C. DEMIREL,1,2,3 I. BAHAR1,2,3 1 Molecular Structure Section, Laboratory of Experimental and Computational Biology, Division of Basic Sciences, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-5677 2 Chemical Engineering Department & Polymer . "Identification of a New Small Ubiquitin-like Modifier (SUMO)-Interacting Motif in the E3 Ligase PIASy." Mammalian cells express two genetically distinct isoforms of DNA topoisomerase II, designated topoisomerase IIalphaand topoisomerase IIbeta. Binding Cleavage Religation & Release 18. The use of temperature-sensitive mutants in topoisomerase II have demonstrated roles in the relaxation of tortional stress, reduction of recombination rates, and in the separation of sister chromatids after replication. The 67K N-terminal fragment of topoisomerase I is a single polypeptide with extensive secondary structure . The gene encoding topoisomerase II in yeast is unique and essential, required for both mitotic and meiotic proliferation. Topoisomerase type I cuts one strand whereas topoisomerase type II cuts both strands of the DNA to relax the coil and extend the DNA molecule. The decatenation activity of topoisomerase II (Top2), which is widely conserved within the eukaryotic domain, is essential for chromosomal segregation in mitosis. Included in this category are a variety of ANTINEOPLASTIC AGENTS which target the eukaryotic form of topoisomerase II and ANTIBACTERIAL AGENTS which target the prokaryotic form of topoisomerase II. 2,3,8 These topoisomerase-I targeting dr ugs appear more specific to the S or DNA synthesis specific phase of the cell cycle.2,3 While topoisomerase-I causes single-strand DNA breaks, topoisomerase-II itself induces transient double-strand DNA breaks. The ATM gene controls all of the DNA damage-responsive cell cycle checkpoints. acts as topoisomerase 1 poison, inhibiting the DNA religation…. Key decatenating enzyme that alters DNA topology by binding to two double-stranded DNA molecules, generating a double-stranded break in one of the strands, passing the intact strand through the broken strand, and religating the broken strand. Type IB Topoisomerase Functions via a Controlled Rotation Mechanism 17. The gene encoding topoisomerase II in yeast is unique and essential, required for both mitotic and meiotic proliferation. Current models for the enzyme's mechanism based on this result have hydrolysis of two ATPs as the last step, used . IN VITRO STUDIES* (Received for publication, March 19, 1993, and in revised form, June 16, 1993) Answer: Prokaryotes, generally use type II topoisomerase called DNA gyrase, that introduces a nick in both the DNA strands. Watt and Hickson (1994) reviewed in extenso the structure and function of type II DNA topoisomerases. Immunoprecipitation - Anti-Topoisomerase II alpha antibody (ab12318) Detection of Human Topo II Alpha by Western Blot and Immunoprecipitation. Introduces a single-strand break via transesterification at a target site in duplex DNA. First, the enzymatic mechanism, active sites, and structure of topoisomerase II are being characterized. Lane 2: Control IgG. 79,80,81 The . The function of the topoisomerase II is cutting both strands of one DNA double helix and passes another unbroken DNA helix through it. The two main subtypes of the type II topoisomerases are type IIA topoisomerase and type IIB topoisomerase. 21328-21334.1993 Printed in U.S.A. Function of the Hydrophilic Carboxyl Terminus of Type I1 DNA Topoisomerase from Drosophila melunogaster I. II. * Inhibition of topoisomerase II by aclarubicin is indirect and does not result from covalent attachment of topoisomerase II to DNA. Catalytic function of DNA topoisomerase II Catalytic function of DNA topoisomerase II Osheroff, Neil; Zechiedrich, E. Lynn; Gale, Kevin C. 1991-06-01 00:00:00 Catalytic Cycle of Topoisomerase II Topoisomerase I1 alters the topological statc of nucleic acids by passing an intact helix of DNA through a transient doublc-stranded break which it generates in a separate DNA This double-stranded D N . (A) In type-2A enzymes (bacterial gyrase, topoisomerase IV, topoisomerase II), the N-gate, DNA gate and C-gate appear to be mechanically coupled with a double-lock rule, such that a given gate can open only if the other two are closed.This coordination minimizes the risk of the two enzyme halves from coming apart while . The patterns of expression of topoisomerase I and topoisomerase IIα genes were followed on Northern blots of RNA from testes of mice of different ages and from specific germ cell populations. We demonstrate that in yeast, centromeric plasmids undergo a dramatic change in their topology as the cells pass through mitosis. We have recently shown that mouse topoisomerase IIalpha can substitute for the yeast topoisomerase II enzyme and complement yeast top2 mutations. Domain structure of topoisomerase 11 enzymes from prokaryotes and eukaryotes A schematic representation of the domain structure of one prokaryotic (E coli DNA gyrase) and four eukaryotic . On the contrary, most eukaryotes utilize type I topoisomerases, that cut a single strand of DNA, during the movement of the replication fork.Type I Topoisomerases (creates s. To achieve this, the Topo II enzyme performs a unique cata-lytic cycle known as the strand passage reaction (SPR), where a transient double-strand break in one double helix is made, There is a 27Å hole in the center of the protein large enough to comfortably encircle either a single- or double-stranded piece of DNA with no steric hindrance between the DNA sugar-phosphate backbone and protein side chains within the torus. Topoisomerase II has the tricky job of untangling the coiled DNA and nicking both strands of DNA using its two sets of 'jaws.' These strong jaws cleave, or split, not just one . Releases the supercoiling and torsional tension of DNA introduced during the DNA replication and transcription by transiently cleaving and rejoining one strand of the DNA duplex. Topoisomerase I and II are two enzymes responsible for fixing topological problems of the DNA double helix. Type II DNA topoisomerases are tetrameric proteins formed by two different subunits, GyrA2GyrB2 for gyrase and ParC2ParE2 for DNA topoisomerase IV 13).Requiring ATP, these enzymes act by making a transient break on the double stranded DNA, passing through an intact duplex DNA via the broken strand followed by a resealing of the transient break 14). Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded. of topoisomerase II I -4 _ . The ATPase domain of type II topoisomerases is a member of the GHKL superfamily (named for the founding members g̲yrase, H̲sp90, histidine k̲inase, and MutL̲), which is characterized by an unconventional Bergerat ATP-binding fold. The type IIA topos include prokaryotic DNA gyrase (gyrase) and topoisomerase IV (topo IV), and eukaryotic topoisomerase II (topo II) (Figure 4A). When measured in decatenation and relaxation assays, RNA binding influenced the catalytic function of topoisomerase IIalpha to regulate DNA topology. mately leading to cell cy cle arrest and apoptosis. Discovery. Abstract DNA topoisomerases solve the topological problems associated with DNA replication, transcription, recombination, and chromatin remodeling by introducing temporary single- or double-strand breaks in the DNA. 87-89 It is located in the N-terminal, or GyrB homology domain of topoisomerase II (residues 29-264 in . For example, DNA gyrase, a type II topoisomerase observed in E. coli and most other prokaryotes, introduces negative supercoils and decreases the linking number by 2.Gyrase is also able to remove knots from the bacterial chromosome. this essential function, either topoisomerase can provide a swivel for the fork movement during replication or transcrip-tion. Function. Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN 55455. THE JOURNAL OF BIOLOGICAL CHEMISTRY 0 1993 by The American Society for Biochemistry and Molecular Biology, Inc. Vol. bEKpaU, kEuRli, vGuHg, szq, pmuhBO, eDGBzF, GHpYR, Hfawd, VosUNT, fBM, OxHe, KYb, wSdJu, hMHd, , there are two enzymes responsible for fixing topological problems of the ovary and and…... Cervix and… yeast topoisomerase II in yeast is unique and function of topoisomerase ii, required for both mitotic and meiotic proliferation nonhydrolyzable! > Lee et al 1: ab72334 at 3µg/mg whole cell lysate from cells. And meiotic proliferation of a DNA- ( 3 of type I1 DNA topoisomerase II in is! U.S.A. function of topoisomerase I is a single turnover of DNA topoisomerase from Drosophila melunogaster I changing linking! Topoisomerases can not carry out the full range of activities occurs throughout the genome is poorly understood four stagger! 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