DOCSLIB.ORG

Transcription-Induced DNA Double Strand Breaks: Both Oncogenic Force and Potential Therapeutic Target?

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Transcription-Induced DNA Double Strand Breaks: Both Oncogenic Force and Potential Therapeutic Target? Published OnlineFirst March 8, 2011; DOI: 10.1158/1078-0432.CCR-10-2044 Clinical Cancer Molecular Pathways Research Transcription-Induced DNA Double Strand Breaks: Both Oncogenic Force and Potential Therapeutic Target? Michael C. Haffner, Angelo M. De Marzo, Alan K. Meeker, William G. Nelson, and Srinivasan Yegnasubramanian Abstract An emerging model of transcriptional activation suggests that induction of transcriptional programs, for instance by stimulating prostate or breast cells with androgens or estrogens, respectively, involves the formation of DNA damage, including DNA double strand breaks (DSB), recruitment of DSB repair proteins, and movement of newly activated genes to transcription hubs. The DSB can be mediated by the class II topoisomerase TOP2B, which is recruited with the androgen receptor and estrogen receptor to regulatory sites on target genes and is apparently required for efficient transcriptional activation of these genes. These DSBs are recognized by the DNA repair machinery triggering the recruitment of repair proteins such as poly(ADP-ribose) polymerase 1 (PARP1), ATM, and DNA-dependent protein kinase (DNA-PK). If illegitimately repaired, such DSBs can seed the formation of genomic rearrangements like the TMPRSS2- ERG fusion oncogene in prostate cancer. Here, we hypothesize that these transcription-induced, TOP2B- mediated DSBs can also be exploited therapeutically and propose that, in hormone-dependent tumors like breast and prostate cancers, a hormone-cycling therapy, in combination with topoisomerase II poisons or inhibitors of the DNA repair components PARP1 and DNA-PK, could overwhelm cancer cells with transcription-associated DSBs. Such strategies may find particular utility in cancers, like prostate cancer, which show low proliferation rates, in which other chemotherapeutic strategies that target rapidly proliferating cells have had limited success. Clin Cancer Res; 17(12); 1–7. Ó2011 AACR. Background damage and repair (5–12). For instance, unbiased proteo- mics approaches as well as candidate protein analyses have The prevailing reductionist view of transcription has shown that proteins such as Ku70, Ku80, poly(ADP-ribose) been challenged by mounting recent evidence suggesting polymerase 1 (PARP1), DNA-dependent protein kinase that induction of transcription involves profound and (DNA-PK), and class II topoisomerase (TOP2B) can be dynamic changes in genome organization and structure. associated with transcription factors including androgen In this emerging view, hundreds of transcription factors receptor (AR) and upstream stimulating factor 1 and can and cofactors are present at locally high concentrations in bind to regulatory elements of target genes upon activation "transcription factories" in the nucleus (1). Induction of of transcription (9, 12). Inhibition of the catalytic function widespread transcriptional changes, for example during of several of these proteins results in an impaired induction hormonal stimulation of hormone-responsive cells, can of transcriptional programs, suggesting a crucial role for result in large-scale alterations in genome organization DNA repair in transcriptional regulation. Among these, arising from movement of different target gene loci and TOP2B is especially interesting because it has been shown distal regulatory elements to these transcriptional hubs, by numerous recent reports to be associated with multiple establishing cell type–specific expression patterns (1–4). nuclear hormone receptors and transcription factors and In addition, recent reports have shown that efficient has also been shown to be bound to promoters and induction of transcriptional programs by a wide array of enhancers of newly transcribed genes (5, 6, 11, 12). Inter- nuclear hormone receptors and other transcription factors estingly, TOP2B catalytic activity appears to be required for involves the recruitment of several factors involved in DNA efficient transcriptional initiation and may be involved in the establishment of complex chromosomal conforma- tional changes during activation of transcription (5, 6, Authors' Affiliation: Sidney Kimmel Comprehensive Cancer Center, 12, 13). Johns Hopkins University School of Medicine, Baltimore, Maryland The two human class II topoisomerases, TOP2A and Corresponding Author: Srinivasan Yegnasubramanian, Johns Hopkins University School of Medicine, 1650 Orleans Street, CRB2, Rm 145, TOP2B, can resolve DNA topologic constraints by passing Baltimore, MD 21231. Phone: 410-502-3425; Fax: 410-502-9817; E-mail: one DNA segment through a transient DNA double strand [email protected] break (DSB) in another segment, in an ATP-dependent doi: 10.1158/1078-0432.CCR-10-2044 fashion (14). In addition to being able to resolve super- Ó2011 American Association for Cancer Research. helical tension introduced by under- or overwinding of the www.aacrjournals.org 1 Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2011 American Association for Cancer Research. Published OnlineFirst March 8, 2011; DOI: 10.1158/1078-0432.CCR-10-2044 Haffner et al AR DHT DNA double strand break DHT AR TOP2B AR DHT TOP2B DHT AR POLII Ku70 Ku80 DNA-PK PARP1 ATM Other Repair Proteins?? ROS Fork collision Dietary carcinogens Repair, movement to Repair infidelity transcription hubs, and active transcription Persistent DSBs Senescence Illegitimate repair Apoptosis Formation of structural rearrangements Genomic instability © 2011 American Association for Cancer Research Figure 1. An emerging model suggests that efficient induction of transcriptional programs can involve the generation of DSBs and perhaps other DNA damage, repair of these breaks via recruitment of DNA repair proteins, and large-scale movement of genes and regulatory elements to transcriptional hubs, in which transcription (co)factors are present at high local concentration. These events are illustrated for induction of transcriptional programs by AR. Upon transcriptional activation, AR associates with TOP2B and binds to AR target sites. DNA repair proteins including Ku70, Ku80, PARP1, ATM, DNA-PK, and possibly other DNA repair enzymes also get recruited to these sites. TOP2B can mediate DSBs in associated DNA. Reactive oxygen species (ROS), perhaps generated during AR signaling, dietary carcinogens, and collision with the replication or transcription forks can transform these transient breaks into frank DSBs. Persistence of these breaks can result in induction of cellular senescence and apoptosis. Illegitimate repair of these breaks can lead to the formation of structural rearrangements. Such pathways are likely at work for other induced transcriptional programs, including those mediated by ER, RAR, and AP1. DNA double helix, the class II topoisomerases are uniquely that TOP2B may be involved in resolving topologic con- able to resolve higher order topologic constraints, such as straints arising from movement of chromosomal segments tangles and knots. TOP2A and TOP2B are encoded by during transcriptional induction. Such a role for TOP2B in separate genes, and although they show a high degree of transcriptional initiation would be analogous to the sequence homology, they differ significantly in their expres- requirement of TOP2A in resolving topologic constraints sion patterns and cellular functions (14, 15). Whereas associated with DNA replication and chromosomal segre- TOP2A is almost exclusively expressed in proliferating cells, gation. where it is required for resolution of topologic constraints Using an elegant assay that allows site-specific labeling of arising during replication and chromosomal segregation, DSBs, Ju and colleagues showed that, during transcriptional TOP2B can be expressed throughout the cell cycle in divid- activation by the estrogen receptor (ER), transient DSBs are ing and nondividing cells (14, 16, 17). Given the recent generated at regulatory elements of ER-regulated genes (6). findings of its association with transcription factors and its This finding was further independently corroborated for catalytic activity at gene regulatory sites, we can speculate other signaling pathways including the AR (Fig. 1; refs. 5, 2 Clin Cancer Res; 17(12) June 15, 2011 Clinical Cancer Research Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2011 American Association for Cancer Research. Published OnlineFirst March 8, 2011; DOI: 10.1158/1078-0432.CCR-10-2044 Targeting Transcription-Induced DSBs for Cancer Therapy 6, 12). Induction of these transient DSBs, tracked with the Regardless of their role in transcriptional regulation, it is recruitment of TOP2B to these sites and targeted depletion of clear that the generation of DNA damage, particularly the TOP2B, dramatically reduced DSB generation, suggesting generation of DSBs, during induction of transcriptional that these transcription-associated DSBs were mediated by programs, may represent a critical vulnerability for cells. TOP2B. Interestingly, these DSBs were recognized by the DSB DSBs play a major role in carcinogenesis: On one hand, repair machinery. The DSB recognition proteins Ku70 and unresolved DSBs can lead to cell cycle arrest, senescence, Ku80, for instance, have been shown to bind together with and apoptosis; and on the other hand, if illegitimately nuclear hormone receptors at target sites (6, 8–10, 12). repaired, such transcription-induced DSBs can seed the Furthermore, the DSB repair–associated protein PARP1 formation of genomic rearrangements, amplifications, and the kinases DNA-PK and ATM were also found
Load more

Recommended publications
  • Targeting Topoisomerase I in the Era of Precision Medicine Anish Thomas and Yves Pommier
    Published OnlineFirst June 21, 2019; DOI: 10.1158/1078-0432.CCR-19-1089 Review Clinical Cancer Research Targeting Topoisomerase I in the Era of Precision Medicine Anish Thomas and Yves Pommier Abstract Irinotecan and topotecan have been widely used as including the indenoisoquinolines LMP400 (indotecan), anticancer drugs for the past 20 years. Because of their LMP776 (indimitecan), and LMP744, and on tumor- selectivity as topoisomerase I (TOP1) inhibitors that trap targeted delivery TOP1 inhibitors using liposome, PEGyla- TOP1 cleavage complexes, camptothecins are also widely tion, and antibody–drug conjugates. We also address how used to elucidate the DNA repair pathways associated with tumor-specific determinants such as homologous recombi- DNA–protein cross-links and replication stress. This review nation defects (HRD and BRCAness) and Schlafen 11 summarizes the basic molecular mechanisms of action (SLFN11) expression can be used to guide clinical appli- of TOP1 inhibitors, their current use, and limitations cation of TOP1 inhibitors in combination with DNA dam- as anticancer agents. We introduce new therapeutic strate- age response inhibitors including PARP, ATR, CHEK1, and gies based on novel TOP1 inhibitor chemical scaffolds ATM inhibitors. Introduction DNA structures such as plectonemes, guanosine quartets, R-loops, and DNA breaks (reviewed in ref. 1). Humans encodes six topoisomerases, TOP1, TOP1MT, TOP2a, TOP2b, TOP3a, and TOP3b (1) to pack and unpack the approx- imately 2 meters of DNA that needs to be contained in the nucleus Anticancer TOP1 Inhibitors Trap TOP1CCs whose diameter (6 mm) is approximately 3 million times smaller. as Interfacial Inhibitors Moreover, the genome is organized in chromosome loops and the separation of the two strands of DNA during transcription and The plant alkaloid camptothecin and its clinical derivatives, replication generate torsional stress and supercoils that are topotecan and irinotecan (Fig.
    [Show full text]
  • Single-Nuclei RNA-Seq on Human Retinal Tissue Provides Improved Transcriptome Profiling
    ARTICLE https://doi.org/10.1038/s41467-019-12917-9 OPEN Single-nuclei RNA-seq on human retinal tissue provides improved transcriptome profiling Qingnan Liang 1,2,3,9, Rachayata Dharmat1,2,8,9, Leah Owen4, Akbar Shakoor4, Yumei Li1, Sangbae Kim1, Albert Vitale4, Ivana Kim4, Denise Morgan4,5, Shaoheng Liang 6, Nathaniel Wu1, Ken Chen 6, Margaret M. DeAngelis4,5,7* & Rui Chen1,2,3* Single-cell RNA-seq is a powerful tool in decoding the heterogeneity in complex tissues by 1234567890():,; generating transcriptomic profiles of the individual cell. Here, we report a single-nuclei RNA- seq (snRNA-seq) transcriptomic study on human retinal tissue, which is composed of mul- tiple cell types with distinct functions. Six samples from three healthy donors are profiled and high-quality RNA-seq data is obtained for 5873 single nuclei. All major retinal cell types are observed and marker genes for each cell type are identified. The gene expression of the macular and peripheral retina is compared to each other at cell-type level. Furthermore, our dataset shows an improved power for prioritizing genes associated with human retinal dis- eases compared to both mouse single-cell RNA-seq and human bulk RNA-seq results. In conclusion, we demonstrate that obtaining single cell transcriptomes from human frozen tissues can provide insight missed by either human bulk RNA-seq or animal models. 1 HGSC, Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA. 2 Department of Molecular and Human Genetics, Baylor College of Medicine, Houston 77030 TX, USA. 3 Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
    [Show full text]
  • The Role of Sumoylation of DNA Topoisomerase Iiα C-Terminal Domain in the Regulation of Mitotic Kinases In
    SUMOylation at the centromere: The role of SUMOylation of DNA topoisomerase IIα C-terminal domain in the regulation of mitotic kinases in cell cycle progression. By Makoto Michael Yoshida Submitted to the graduate degree program in the Department of Molecular Biosciences and the Graduate Faculty of the University of Kansas in partial fulfillment of the requirements for the degree of Doctor of Philosophy. ________________________________________ Chairperson: Yoshiaki Azuma, Ph.D. ________________________________________ Roberto De Guzman, Ph.D. ________________________________________ Kristi Neufeld, Ph.D. _________________________________________ Berl Oakley, Ph.D. _________________________________________ Blake Peterson, Ph.D. Date Defended: July 12, 2016 The Dissertation Committee for Makoto Michael Yoshida certifies that this is the approved version of the following dissertation: SUMOylation at the centromere: The role of SUMOylation of DNA topoisomerase IIα C-terminal domain in the regulation of mitotic kinases in cell cycle progression. ________________________________________ Chairperson: Yoshiaki Azuma, Ph.D. Date approved: July 12, 2016 ii ABSTRACT In many model systems, SUMOylation is required for proper mitosis; in particular, chromosome segregation during anaphase. It was previously shown that interruption of SUMOylation through the addition of the dominant negative E2 SUMO conjugating enzyme Ubc9 in mitosis causes abnormal chromosome segregation in Xenopus laevis egg extract (XEE) cell-free assays, and DNA topoisomerase IIα (TOP2A) was identified as a substrate for SUMOylation at the mitotic centromeres. TOP2A is SUMOylated at K660 and multiple sites in the C-terminal domain (CTD). We sought to understand the role of TOP2A SUMOylation at the mitotic centromeres by identifying specific binding proteins for SUMOylated TOP2A CTD. Through affinity isolation, we have identified Haspin, a histone H3 threonine 3 (H3T3) kinase, as a SUMOylated TOP2A CTD binding protein.
    [Show full text]
  • Differential Expression of Multiple Disease-Related Protein Groups
    brain sciences Article Differential Expression of Multiple Disease-Related Protein Groups Induced by Valproic Acid in Human SH-SY5Y Neuroblastoma Cells 1,2, 1, 1 1 Tsung-Ming Hu y, Hsiang-Sheng Chung y, Lieh-Yung Ping , Shih-Hsin Hsu , Hsin-Yao Tsai 1, Shaw-Ji Chen 3,4 and Min-Chih Cheng 1,* 1 Department of Psychiatry, Yuli Branch, Taipei Veterans General Hospital, Hualien 98142, Taiwan; [email protected] (T.-M.H.); [email protected] (H.-S.C.); [email protected] (L.-Y.P.); fi[email protected] (S.-H.H.); [email protected] (H.-Y.T.) 2 Department of Future Studies and LOHAS Industry, Fo Guang University, Jiaosi, Yilan County 26247, Taiwan 3 Department of Psychiatry, Mackay Medical College, New Taipei City 25245, Taiwan; [email protected] 4 Department of Psychiatry, Taitung Mackay Memorial Hospital, Taitung County 95064, Taiwan * Correspondence: [email protected]; Tel.: +886-3888-3141 (ext. 475) These authors contributed equally to this work. y Received: 10 July 2020; Accepted: 8 August 2020; Published: 12 August 2020 Abstract: Valproic acid (VPA) is a multifunctional medication used for the treatment of epilepsy, mania associated with bipolar disorder, and migraine. The pharmacological effects of VPA involve a variety of neurotransmitter and cell signaling systems, but the molecular mechanisms underlying its clinical efficacy is to date largely unknown. In this study, we used the isobaric tags for relative and absolute quantitation shotgun proteomic analysis to screen differentially expressed proteins in VPA-treated SH-SY5Y cells. We identified changes in the expression levels of multiple proteins involved in Alzheimer’s disease, Parkinson’s disease, chromatin remodeling, controlling gene expression via the vitamin D receptor, ribosome biogenesis, ubiquitin-mediated proteolysis, and the mitochondrial oxidative phosphorylation and electron transport chain.
    [Show full text]
  • Modification of Topoisomerase Genes Copy Number in Newly Diagnosed
    Leukemia (2003) 17, 532–540 & 2003 Nature Publishing Group All rights reserved 0887-6924/03 $25.00 www.nature.com/leu Modification of topoisomerase genes copy number in newly diagnosed childhood acute lymphoblastic leukemia E Gue´rin1,2, N Entz-Werle´3, D Eyer3, E Pencreac’h1, A Schneider1, A Falkenrodt4, F Uettwiller3, A Babin3, A-C Voegeli1,5, M Lessard4, M-P Gaub1,2, P Lutz3 and P Oudet1,5 1Laboratoire de Biochimie et de Biologie Mole´culaire Hoˆpital de Hautepierre, Strasbourg, France; 2INSERM U381, Strasbourg, France; 3Service d’Onco-He´matologie Pe´diatrique Hoˆpital de Hautepierre, Strasbourg, France; 4Laboratoire Hospitalier d’He´matologie Biologique Hoˆpital de Hautepierre, Strasbourg, France; and 5INSERM U184, Illkirch, France Topoisomerase genes were analyzed at both DNA and RNA segregation.4 These enzymes act by promoting transient DNA levels in 25 cases of newly diagnosed childhood acute breakage in order to allow strand-passage events and DNA lymphoblastic leukemia (ALL). The results of molecular analy- 5 sis were compared to risk group classification of children in relaxation to occur before rejoining the broken DNA ends. order to identify molecular characteristics associated with Two types of DNA topoisomerases have been described. Type response to therapy. At diagnosis, allelic imbalance at topo- I enzymes, encoded in humans by the TOP1, TOP3A and isomerase IIa (TOP2A) gene locus was found in 75% of TOP3B genes,6–8 cleave only one strand of the DNA helix informative cases whereas topoisomerase I and IIb gene loci whereas type II enzymes, encoded by the human TOP2A and are altered in none or only one case, respectively.
    [Show full text]
  • Cyclin a Triggers Mitosis Either Via Greatwall Or Cyclin B
    bioRxiv preprint doi: https://doi.org/10.1101/501684; this version posted December 20, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Cyclin A triggers Mitosis either via Greatwall or Cyclin B Nadia Hégarat(1)*, Adrijana Crncec(1)*, Maria F. Suarez Peredoa Rodri- guez(1), Fabio Echegaray Iturra(1), Yan Gu(1), Paul F. Lang(2), Alexis R. Barr(3), Chris Bakal(4), Masato T. Kanemaki(5), Angus I. Lamond(6), Bela Novak(2), Tony Ly(7)•• and Helfrid Hochegger(1)•• (1) Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN19RQ, UK (2) Department of Biochemistry, University of Oxford, South Park Road, Oxford OX13QU, UK (3) MRC London Institute of Medical Science, Imperial College, London W12 0NN, UK (4) The Institute of Cancer Research, London SW3 6JB, UK (5) National Institute of Genetics, Research Organization of Information and Sys- tems (ROIS), and Department of Genetics, SOKENDAI (The Graduate University of Advanced Studies), Yata 1111, Mishima, Shizuoka 411-8540, Japan. (6) Centre for Gene Regulation and Expression, School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK (7) Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3BF, UK * Equal contribution ** Correspondence: Tony Ly: [email protected]; Helfrid Hochegger: [email protected] bioRxiv preprint doi: https://doi.org/10.1101/501684; this version posted December 20, 2018.
    [Show full text]
  • TOP2B)Ina Patient with Acute Myeloid Leukemia with a New T(3;11)(P24;P15) Karin Nebral,1Helmut H
    Human Cancer Biology NUP98 Is Fused to Topoisomerase (DNA) IIB 18 0 kDa (TOP2B)ina Patient with Acute Myeloid Leukemia with a New t(3;11)(p24;p15) Karin Nebral,1Helmut H. Schmidt,2 Oskar A. Haas,1and Sabine Strehl1 Abstract Purpose: The nucleoporin 98 kDa (NUP98) gene has been reported to be fused to 17 different partner genes in various hematologic malignancies with11p15aberrations. Cytogenetic analysis of an adult de novo acute myelogenous leukemia (M5a) revealed a t(3;11)(p24;p15), suggesting rearrangement of NUP98 with a novel partner gene. Experimental Design: Fluorescence in situ hybridization (FISH) was used to confirm the involvement of NUP98 in the t(3;11)(p24;p15). Selection of possibleNUP98 partner genes was done by computer-aided analysis of the 3p24 region using the University of California Santa Cruz genome browser. Fusion gene ^ specific FISH and reverse transcription-PCR analyses were done to verify the presence of the new NUP98 fusion. Results: FISH analysis using a NUP98 -specific clone showed a split signal, indicating that the NUP98 gene was affected by the translocation. Of the genes localized at 3p24, TOP2B was selected as a possible fusion partner candidate gene. Dual-color fusion gene ^ specific FISH and reverse transcription-PCR analysis verified that NUP98 was indeed fused to TOP2B.Inaddition to reciprocal NUP98-TOP2B and TOP2B-NUP98 in-frame fusion transcripts, an alternatively spliced out-of-frame TOP2B-NUP98 transcript that resulted in a premature stop codon was detected. Analysis of the genomic breakpoints revealed typical signs of nonhomologous end joining resulting from error-prone DNA repair.
    [Show full text]
  • How Does SUMO Participate in Spindle Organization?
    cells Review How Does SUMO Participate in Spindle Organization? Ariane Abrieu * and Dimitris Liakopoulos * CRBM, CNRS UMR5237, Université de Montpellier, 1919 route de Mende, 34090 Montpellier, France * Correspondence: [email protected] (A.A.); [email protected] (D.L.) Received: 5 July 2019; Accepted: 30 July 2019; Published: 31 July 2019 Abstract: The ubiquitin-like protein SUMO is a regulator involved in most cellular mechanisms. Recent studies have discovered new modes of function for this protein. Of particular interest is the ability of SUMO to organize proteins in larger assemblies, as well as the role of SUMO-dependent ubiquitylation in their disassembly. These mechanisms have been largely described in the context of DNA repair, transcriptional regulation, or signaling, while much less is known on how SUMO facilitates organization of microtubule-dependent processes during mitosis. Remarkably however, SUMO has been known for a long time to modify kinetochore proteins, while more recently, extensive proteomic screens have identified a large number of microtubule- and spindle-associated proteins that are SUMOylated. The aim of this review is to focus on the possible role of SUMOylation in organization of the spindle and kinetochore complexes. We summarize mitotic and microtubule/spindle-associated proteins that have been identified as SUMO conjugates and present examples regarding their regulation by SUMO. Moreover, we discuss the possible contribution of SUMOylation in organization of larger protein assemblies on the spindle, as well as the role of SUMO-targeted ubiquitylation in control of kinetochore assembly and function. Finally, we propose future directions regarding the study of SUMOylation in regulation of spindle organization and examine the potential of SUMO and SUMO-mediated degradation as target for antimitotic-based therapies.
    [Show full text]
  • The Deubiquitinating Enzyme Inhibitor PR-619 Is a Potent DNA Topoisomerase II Poison
    Molecular Pharmacology Fast Forward. Published on September 12, 2019 as DOI: 10.1124/mol.119.117390 This article has not been copyedited and formatted. The final version may differ from this version. MOL # 117390 The deubiquitinating enzyme inhibitor PR-619 is a potent DNA topoisomerase II poison Ian G. Cowell, Elise M. Ling, Rebecca L. Swan, Matilda L. W. Brooks, & Caroline A. Austin Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne. NE2 4HH. U.K. (I.G.C., E.M.L, R.L.S., M.L.W.B,. & C.A.A.) Downloaded from molpharm.aspetjournals.org at ASPET Journals on September 27, 2021 1 Molecular Pharmacology Fast Forward. Published on September 12, 2019 as DOI: 10.1124/mol.119.117390 This article has not been copyedited and formatted. The final version may differ from this version. MOL # 117390 Running title: PR-619 is a TOP2 poison Corresponding authors To whom correspondence should be addressed: Dr Ian G. Cowell and Professor Caroline A. Austin Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne NE2 4HH. United Kingdom Tel: +44 (0)191 208 7677 Fax: +44 (0)191 208 7424 Email: [email protected] Downloaded from Manuscript information 24 text pages including references and Figure legends 8 figures molpharm.aspetjournals.org 0 tables 48 number of references 185 words in abstract 586 words in Introduction 920 words in Discussion Non standard abbreviations at ASPET Journals on September 27, 2021 DMSO, Dimethyl sulfoxide; DSB, DNA double-strand break; DUB, deubiquitinating enzyme inhibitor; H2AX, histone H2A.X; γH2AX, S-139 phospho-histone H2A.X; TOP2A, DNA Topoisomerase IIα; TOP2B, DNA topoisomerase IIβ; TOP2, DNA topoisomerase II.
    [Show full text]
  • Identification of Novel Genes in BRCA1-Regulated Pathways
    Identification of Novel Genes in BRCA1-Regulated Pathways DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Shweta Kotian, M.S. Graduate Program in Molecular, Cellular and Developmental Biology The Ohio State University 2013 Dissertation Committee: Professor Jeffrey D. Parvin, Advisor Professor Joanna L. Groden Professor Denis C. Guttridge Professor Mark R. Parthun Copyright by Shweta Kotian 2013 Abstract BRCA1 is an important breast- and ovarian-specific tumor suppressor gene. It is important in various cellular functions in the body including transcriptional regulation, cell cycle checkpoint activation, DNA damage response, and maintenance of genomic stability. Approximately 40% of hereditary breast cancers have mutations in BRCA1 or BRCA2, and it is unclear how the remaining cases are caused, presumably by mutations or in the alteration of expression of unknown genes. We hypothesize that these unknown genes or ‘missing BRCAs’ can be identified by bio-informatics methods. Performing gene co-expression analysis, we have compared the expression profiles of hundreds of genes across publicly available breast cancer microarray datasets, with those of BRCA1 and BRCA2. We propose that the genes, whose expression is highly correlated with BRCA1 and BRCA2, might function together in the same pathways. They could also potentially interact with each other. The main aim of this study is to identify these genes, and then biologically validate them in functional BRCA1-regulated pathways including homologous recombination and centrosome duplication. We also hope to find any mechanistic ii associations between these genes and BRCA1. Eventually, we hope to evaluate if these genes contribute to the process of carcinogenesis.
    [Show full text]
  • Type II DNA Topoisomerases Cause Spontaneous Double-Strand Breaks in Genomic DNA
    G C A T T A C G G C A T genes Review Type II DNA Topoisomerases Cause Spontaneous Double-Strand Breaks in Genomic DNA Suguru Morimoto 1, Masataka Tsuda 1, Heeyoun Bunch 2 , Hiroyuki Sasanuma 1, Caroline Austin 3 and Shunichi Takeda 1,* 1 Department of Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan; [email protected] (S.M.); [email protected] (M.T.); [email protected] (H.S.) 2 Department of Applied Biosciences, College of Agriculture and Life Sciences, Kyungpook National University, Daegu 41566, Korea; [email protected] 3 The Institute for Cell and Molecular Biosciences, the Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne NE2 4HH, UK; [email protected] * Correspondence: [email protected]; Tel.: +81-(075)-753-4411 Received: 30 September 2019; Accepted: 26 October 2019; Published: 30 October 2019 Abstract: Type II DNA topoisomerase enzymes (TOP2) catalyze topological changes by strand passage reactions. They involve passing one intact double stranded DNA duplex through a transient enzyme-bridged break in another (gated helix) followed by ligation of the break by TOP2. A TOP2 poison, etoposide blocks TOP2 catalysis at the ligation step of the enzyme-bridged break, increasing the number of stable TOP2 cleavage complexes (TOP2ccs). Remarkably, such pathological TOP2ccs are formed during the normal cell cycle as well as in postmitotic cells. Thus, this ‘abortive catalysis’ can be a major source of spontaneously arising DNA double-strand breaks (DSBs).
    [Show full text]
  • Distinct Expression Patterns for Type II Topoisomerases IIA and IIB in the Early Foetal Human Telencephalon
    Harkin LF, Gerrelli D, Diaz DCG, Santos C, Alzu'bi A, Austin CA, Clowry GJ. Distinct expression patterns for type II topoisomerases IIA and IIB in the early foetal human telencephalon. Journal of Anatomy 2015, DOI: 10.1111/joa.12416 Copyright: © 2015 The Authors. Journal of Anatomy published by John Wiley & Sons Ltd on behalf of Anatomical Society. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. DOI link to article: http://dx.doi.org/10.1111/joa.12416 Date deposited: 18/12/2015 This work is licensed under a Creative Commons Attribution 4.0 International License Newcastle University ePrints - eprint.ncl.ac.uk Journal of Anatomy J. Anat. (2015) doi: 10.1111/joa.12416 Distinct expression patterns for type II topoisomerases IIA and IIB in the early foetal human telencephalon Lauren F. Harkin,1,2 Dianne Gerrelli,3 Diana C. Gold Diaz,3 Chloe Santos,3 Ayman Alzu’bi,1,2 Caroline A. Austin4 and Gavin J. Clowry1 1Institute of Neuroscience, Newcastle University, Newcastle upon Tyne, UK 2Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK 3HDBR Resource, UCL Institute of Child Health, London, UK 4Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne, UK Abstract TOP2A and TOP2B are type II topoisomerase enzymes that have important but distinct roles in DNA replication and RNA transcription. Recently, TOP2B has been implicated in the transcription of long genes in particular that play crucial roles in neural development and are susceptible to mutations contributing to neurodevelopmental conditions such as autism and schizophrenia.
    [Show full text]