DOCSLIB.ORG

DNA Distress: Just Ring 9-1-1 [9,11]

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
DNA Distress: Just Ring 9-1-1 [9,11] CORE Metadata, citation and similar papers at core.ac.uk Provided by Elsevier - Publisher Connector Dispatch R733 DNA Distress: Just Ring 9-1-1 [9,11]. Interestingly, the Rad9 carboxyl terminus plays an important role in activation of the DNA damage The Rad9–Hus1–Rad1 checkpoint clamp (9-1-1) is a central player in the cellular checkpoint, which halts cell cycle response to DNA damage; three groups have determined the crystal structure progression to provide time for of 9-1-1, providing new insight into its loading mechanism and association with DNA repair [1]. Phosphorylation of DNA damage checkpoint and repair enzymes. this domain is required for binding and recruiting TopBP1 to sites of Michael Kemp and Aziz Sancar PCNA and 9-1-1 require the DNA damage [17] in order to activity of heteropentameric clamp activate the DNA damage response The genome is constantly exposed to loading complexes termed replication kinase ATR [18]. Unfortunately, to cellular metabolites and exogenous factor C (RFC) and Rad17-RFC, obtain recombinant 9-1-1 protein agents that induce lesions in DNA respectively, which bind to, open, suitable for crystallization, all capable of causing mutation, cancer, and clamp the complexes around three groups [9–11] used a truncated or cell death. In response to such DNA [13]. Whereas PCNA is loaded form of Rad9 lacking this region, damage, eukaryotic cells activate onto 30-primer–template junctions and hence the structures provide signaling pathways that promote DNA by the canonical RFC made up of no insight into the role of the Rad9 repair, allow bypass of lesions during RFC1–5, 9-1-1 is instead preferentially carboxyl terminus in the checkpoint DNA replication, and halt cell-cycle loaded onto 50-recessed ends by response. progression [1,2]. When damage is an alternative form of RFC containing However, by analyzing the DNA extensive or unrepairable, these the Rad17 protein in place of RFC1 binding abilities of 9-1-1 containing pathways may instead induce cell [14,15]. It is expected that the full-length or truncated Rad9, Sohn death through apoptosis. Though different loading properties of and Cho [9] provide evidence that the many proteins mediate the various PCNA and 9-1-1 may be linked Rad9 carboxy-terminal tail prevents aspects of the DNA damage to their unique roles 9-1-1 from binding to DNA containing response, a key component of this in DNA metabolism. 50-recessed primer-template junctions. network is the heterotrimeric Whereas the three identical subunits Though the mechanism of this complex Rad9–Hus1–Rad1 (9-1-1) of PCNA impart symmetry to its inhibition is not known, the authors (Figure 1). structure and provide the same suggest that the carboxy-terminal Computational modeling [3], electron interaction surface to RFC, the domain may physically block DNA from microscopy (EM) [4,5], and biochemical presence of three unique subunits in entering the central hole in 9-1-1. It analyses [4,6,7] had already revealed 9-1-1 suggests that its loading onto is therefore plausible that the clamp the 9-1-1 complex to be structurally DNA is more tightly regulated. Yeast loader Rad17-RFC or other factors similar to proliferating cell nuclear two-hybrid and pull-down assays that interact with the Rad9 carboxyl antigen (PCNA), a homotrimeric protein showing direct interactions of Rad17 terminus may alleviate its negative complex that encircles DNA and with the Rad1 and Rad9 subunits of effect during the course of 9-1-1 stimulates DNA polymerases and 9-1-1 [4,10,16] implicate these two loading onto DNA. Consistent with other enzymes involved in DNA subunits in clamp opening. Indeed, replication and repair. Though genetic through an examination of subunit and biochemical approaches show interface buried surface areas and largely separate roles for PCNA and computational analyses of interface Apoptosis DNA repair 9-1-1 in DNA metabolism, several shape complementarity, Dore´ et al. proteins functionally interact with [11] and Xu et al. [10] conclude the both complexes, including flap Rad9–Rad1 interface is weakest and endonuclease I (FEN1) and DNA therefore the most probable location Ligase I [8]. With recent reports of for ring opening. Sohn and Cho [9], 9-1-1 crystal structures [9–11],itis however, suggest that the Hus1-Rad1 9-1-1 now possible to make direct and interface may be optimal for opening detailed comparisons with PCNA because, they argue, it appears structure in order to better structurally most similar to the understand the distinct functions subunit interfaces in PCNA. DNA damage Translesion of the two clamps. Additional work is clearly required checkpoints synthesis All three independently obtained to characterize the mechanism of structures show a closed-ring 9-1-1 clamp loading onto DNA by Current Biology architecture for 9-1-1 (Figure 2), formed Rad17–RFC, and may well reveal that through a head-to-tail association of more than one loading mechanism Figure 1. The Rad9-Hus1-Rad1 (9-1-1) clamp its subunits, just as in PCNA [12]. is employed. plays a central role in the cellular response to Consistent with a previous EM An additional difference between DNA damage. analysis of the 9-1-1 complex on DNA 9-1-1 and PCNA is the presence of Through direct interactions with ligand [4], these structures show the central a carboxy-terminal w120 amino acid protein binding partners, the 9-1-1 complex functions in multiple DNA damage response hole in 9-1-1 to contain sufficient tail on Rad9 that computational pathways, including DNA damage check- space for double-stranded DNA analysis and limited proteolysis points, DNA repair, translesion synthesis, (dsDNA). To encircle DNA, both experiments predict to be unstructured and apoptosis. Current Biology Vol 19 No 17 R734 distributions, unlike the IDLs in the Tsurimoto, T. (2002). Clamp and clamp loader structures of the human checkpoint protein homotrimeric PCNA. This suggests complexes, Rad9-1-1 and Rad17-RFC. Genes that the association of ligand proteins Cells 7, 861–868. to 9-1-1 may involve a preferred 6. Burtelow, M.A., Roos-Mattjus, P.M., Rauen, M., Babendure, J.R., and Karnitz, L.M. (2001). subunit, and Xu et al. [10] indeed show Reconstitution and molecular analysis of the that though all three subunits of 9-1-1 hRad9-hHus1-hRad1 (9-1-1) DNA damage responsive checkpoint complex. J. Biol. Chem. are capable of binding a peptide from 276, 25903–25909. FEN1, Rad1 does so with the highest 7. Lindsey-Boltz, L.A., Bermudez, V.P., affinity [10]. Hurwitz, J., and Sancar, A. (2001). Purification and characterization of Consistent with these biochemical human DNA damage checkpoint Rad data, Xu et al. [10] obtained a co-crystal complexes. Proc. Natl. Acad. Sci. USA 98, 11236–11241. of 9-1-1 with a FEN1 peptide that 8. Helt, C.E., Wang, W., Keng, P.C., and shows a direct interaction with Bambara, R.A. (2005). Evidence that DNA Rad1. Interestingly, and unlike PCNA, damage detection machinery participates in DNA repair. Cell Cycle 4, 529–532. Dore´ et al. report that FEN1 does 9. Sohn, S.Y., and Cho, Y. (2009). Crystal structure not require its PIP box to stably of the human rad9-hus1-rad1 clamp. J. Mol. Biol. 390, 490–502. interact with 9-1-1 in pull-down 10. Xu, M., Bai, L., Gong, Y., Xie, W., Hang, H., and Figure 2. Crystal structure of the human assays [11]. Furthermore, whereas Jiang, T. (2009). Structure and functional 9-1-1 complex. the presence of three equivalent implications of the human Rad9-Hus1-Rad1 Atomic coordinates deposited under ID code cell cycle checkpoint complex. J. Biol. Chem. subunits in PCNA allows the binding 284, 20457–20461. 3G65 by Dore´ et al. [11] were obtained from of up to three proteins simultaneously, 11. Dore, A.S., Kilkenny, M.L., Rzechorzek, N.J., the RCSB Protein Data Bank and analyzed and Pearl, L.H. (2009). Crystal structure using the Jmol viewer. binding to 9-1-1 appears to be of the rad9-rad1-hus1 DNA damage exclusive, as FEN1 readily inhibits checkpoint complex–implications for the association of 9-1-1 with a peptide clamp loading and regulation. Mol. Cell 34, 735–745. such a model, recent NMR data show from the cell cycle inhibitor protein 12. Krishna, T.S., Kong, X.P., Gary, S., that the Rad9 carboxy-terminal domain p21 [11]. These results therefore Burgers, P.M., and Kuriyan, J. (1994). Crystal structure of the eukaryotic DNA binds the single-stranded DNA binding indicate distinct differences in polymerase processivity factor PCNA. Cell 79, protein RPA [19], and the stable ligand protein binding mechanism 1233–1243. association of 9-1-1 at sites of between 9-1-1 and PCNA. 13. Majka, J., and Burgers, P.M. (2004). The PCNA-RFC families of DNA clamps and replication stress was recently shown Ever since the discovery of 9-1-1, clamp loaders. Prog. Nucleic Acid Res. to require the presence of TopBP1 [20]. an outstanding question has been if it Mol. Biol. 78, 227–260. 14. Ellison, V., and Stillman, B. (2003). Biochemical Together, these results imply that looks like PCNA and walks like PCNA, characterization of DNA damage checkpoint a series of coordinated interactions why is 9-1-1 needed? The availability complexes: clamp loader and clamp of 9-1-1 and the Rad9 carboxyl of 9-1-1 crystal structures partially complexes with specificity for 50 recessed DNA. PLoS Biol. 1, E33. terminus with Rad17–RFC, TopBP1, addresses this issue, and this 15. Majka, J., Binz, S.K., Wold, M.S., and and RPA may impact the loading structural information should now Burgers, P.M.
Load more

Recommended publications
  • ATR Pathway Inhibition Is Synthetically Lethal in Cancer Cells with ERCC1 Deficiency
    Published OnlineFirst March 24, 2014; DOI: 10.1158/0008-5472.CAN-13-3229 Cancer Therapeutics, Targets, and Chemical Biology Research ATR Pathway Inhibition Is Synthetically Lethal in Cancer Cells with ERCC1 Deficiency Kareem N. Mohni, Gina M. Kavanaugh, and David Cortez Abstract The DNA damage response kinase ATR and its effector kinase CHEK1 are required for cancer cells to survive oncogene-induced replication stress. ATR inhibitors exhibit synthetic lethal interactions, with deficiencies in the DNA damage response enzymes ATM and XRCC1 and with overexpression of the cell cycle kinase cyclin E. Here, we report a systematic screen to identify synthetic lethal interactions with ATR pathway–targeted drugs, rationalized by their predicted therapeutic utility in the oncology clinic. We found that reduced function in the ATR pathway itself provided the strongest synthetic lethal interaction. In addition, we found that loss of the structure-specific endonuclease ERCC1-XPF (ERCC4) is synthetic lethal with ATR pathway inhibitors. ERCC1- deficient cells exhibited elevated levels of DNA damage, which was increased further by ATR inhibition. When treated with ATR or CHEK1 inhibitors, ERCC1-deficient cells were arrested in S-phase and failed to complete cell-cycle transit even after drug removal. Notably, triple-negative breast cancer cells and non–small cell lung cancer cells depleted of ERCC1 exhibited increased sensitivity to ATR pathway–targeted drugs. Overall, we concluded that ATR pathway–targeted drugs may offer particular utility in cancers with reduced ATR pathway function or reduced levels of ERCC4 activity. Cancer Res; 74(10); 1–11. Ó2014 AACR. Introduction repair pathway such as homologous recombination or post- – Replicating DNA is sensitive to a wide array of endogenous replicative repair to remove the PARP DNA complexes (7).
    [Show full text]
  • Complex Interacts with WRN and Is Crucial to Regulate Its Response to Replication Fork Stalling
    Oncogene (2012) 31, 2809–2823 & 2012 Macmillan Publishers Limited All rights reserved 0950-9232/12 www.nature.com/onc ORIGINAL ARTICLE The RAD9–RAD1–HUS1 (9.1.1) complex interacts with WRN and is crucial to regulate its response to replication fork stalling P Pichierri, S Nicolai, L Cignolo1, M Bignami and A Franchitto Department of Environment and Primary Prevention, Istituto Superiore di Sanita`, Rome, Italy The WRN protein belongs to the RecQ family of DNA preference toward substrates that mimic structures helicases and is implicated in replication fork restart, but associated with stalled replication forks (Brosh et al., how its function is regulated remains unknown. We show 2002; Machwe et al., 2006) and WS cells exhibit that WRN interacts with the 9.1.1 complex, one of enhanced instability at common fragile sites, chromo- the central factors of the replication checkpoint. This somal regions especially prone to replication fork interaction is mediated by the binding of the RAD1 stalling (Pirzio et al., 2008). How WRN favors recovery subunit to the N-terminal region of WRN and is of stalled forks and prevents DNA breakage upon instrumental for WRN relocalization in nuclear foci and replication perturbation is not fully understood. It has its phosphorylation in response to replication arrest. We been suggested that WRN might facilitate replication also find that ATR-dependent WRN phosphorylation restart by either promoting recombination or processing depends on TopBP1, which is recruited by the 9.1.1 intermediates at stalled forks in a way that counteracts complex in response to replication arrest. Finally, we unscheduled recombination (Franchitto and Pichierri, provide evidence for a cooperation between WRN and 2004; Pichierri, 2007; Sidorova, 2008).
    [Show full text]
  • Supplementary Table S1. Correlation Between the Mutant P53-Interacting Partners and PTTG3P, PTTG1 and PTTG2, Based on Data from Starbase V3.0 Database
    Supplementary Table S1. Correlation between the mutant p53-interacting partners and PTTG3P, PTTG1 and PTTG2, based on data from StarBase v3.0 database. PTTG3P PTTG1 PTTG2 Gene ID Coefficient-R p-value Coefficient-R p-value Coefficient-R p-value NF-YA ENSG00000001167 −0.077 8.59e-2 −0.210 2.09e-6 −0.122 6.23e-3 NF-YB ENSG00000120837 0.176 7.12e-5 0.227 2.82e-7 0.094 3.59e-2 NF-YC ENSG00000066136 0.124 5.45e-3 0.124 5.40e-3 0.051 2.51e-1 Sp1 ENSG00000185591 −0.014 7.50e-1 −0.201 5.82e-6 −0.072 1.07e-1 Ets-1 ENSG00000134954 −0.096 3.14e-2 −0.257 4.83e-9 0.034 4.46e-1 VDR ENSG00000111424 −0.091 4.10e-2 −0.216 1.03e-6 0.014 7.48e-1 SREBP-2 ENSG00000198911 −0.064 1.53e-1 −0.147 9.27e-4 −0.073 1.01e-1 TopBP1 ENSG00000163781 0.067 1.36e-1 0.051 2.57e-1 −0.020 6.57e-1 Pin1 ENSG00000127445 0.250 1.40e-8 0.571 9.56e-45 0.187 2.52e-5 MRE11 ENSG00000020922 0.063 1.56e-1 −0.007 8.81e-1 −0.024 5.93e-1 PML ENSG00000140464 0.072 1.05e-1 0.217 9.36e-7 0.166 1.85e-4 p63 ENSG00000073282 −0.120 7.04e-3 −0.283 1.08e-10 −0.198 7.71e-6 p73 ENSG00000078900 0.104 2.03e-2 0.258 4.67e-9 0.097 3.02e-2 Supplementary Table S2.
    [Show full text]
  • Structure and Functional Implications of the Human Rad9-Hus1-Rad1
    Supplemental Material can be found at: http://www.jbc.org/content/suppl/2009/06/17/C109.022384.DC1.html ACCELERATED PUBLICATION This paper is available online at www.jbc.org THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 284, NO. 31, pp. 20457–20461, July 31, 2009 © 2009 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in the U.S.A. Structure and Functional onto DNA lesion sites by Rad17-RFC2–5 (which consists of one large subunit, Rad17, and four small subunits, RFC2–5), where Implications of the Human it triggers the activation of the cell cycle checkpoint (3, 4). Rad9-Hus1-Rad1 Cell Cycle Moreover, the 9-1-1 complex can also directly participate in DNA repair via physical association with many factors involved *□S Checkpoint Complex in base excision repair (BER), translesion synthesis, homolo- Received for publication, May 18, 2009, and in revised form, June 5, 2009 gous recombination, and mismatch repair pathways (5–9). Published, JBC Papers in Press, June 17, 2009, DOI 10.1074/jbc.C109.022384 Although both the 9-1-1 and the PCNA complexes perform Min Xu‡§, Lin Bai‡§, Yong Gong‡, Wei Xie‡§, Haiying Hang‡1, ‡2 critical functions in eukaryotic cells with predicted similar and Tao Jiang structures (10), their specific roles are distinct. First, the 9-1-1 ‡ From the National Laboratory of Biomacromolecules, Institute of complex is a DNA damage sensor in the cell cycle checkpoint Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 and the §Graduate University of Chinese Academy but does not function as a scaffold for the major DNA replica- of Sciences, 19A Yuquan Road, Shijingshan District, Beijing 100039, China tion factors; however, PCNA plays exactly the opposite role (1, 11).
    [Show full text]
  • Signature of Progression and Negative Outcome in Colorectal Cancer
    Oncogene (2010) 29, 876–887 & 2010 Macmillan Publishers Limited All rights reserved 0950-9232/10 $32.00 www.nature.com/onc ORIGINAL ARTICLE A ‘DNA replication’ signature of progression and negative outcome in colorectal cancer M-J Pillaire1,7, J Selves2,7, K Gordien2, P-A Gouraud3, C Gentil3, M Danjoux2,CDo3, V Negre4, A Bieth1, R Guimbaud2, D Trouche5, P Pasero6,MMe´chali6, J-S Hoffmann1 and C Cazaux1 1Genetic Instability and Cancer Group, Department Biology of Cancer, Institute of Pharmacology and Structural Biology, UMR5089 CNRS, University of Toulouse, University Paul Sabatier, Toulouse, France; 2INSERM U563, Federation of Digestive Cancerology and Department of Anatomo-pathology, University of Toulouse, University Paul Sabatier, Toulouse, France; 3Service of Epidemiology, INSERM U558, Faculty of Medicine, University of Toulouse, University Paul Sabatier, Alle´es Jules Guesde, Toulouse, France; 4aCGH GSO Canceropole Platform, INSERM U868, Val d’Aurelle, Montpellier, France; 5Laboratory of Cellular and Molecular Biology of Cell Proliferation Control, UMR 5099 CNRS, University of Toulouse, University Paul Sabatier, Toulouse, France and 6Institute of Human Genetics UPR1142 CNRS, Montpellier, France Colorectal cancer is one of the most frequent cancers Introduction worldwide. As the tumor-node-metastasis (TNM) staging classification does not allow to predict the survival of DNA replication in normal cells is regulated by an patients in many cases, additional prognostic factors are ‘origin licensing’ mechanism that ensures that it occurs needed to better forecast their outcome. Genes involved in just once per cycle. Once cells enter the S-phase, the DNA replication may represent an underexplored source stability of DNA replication forks must be preserved to of such prognostic markers.
    [Show full text]
  • A Review of Prostate Cancer Genome-Wide Association Studies (GWAS)
    Published OnlineFirst January 18, 2018; DOI: 10.1158/1055-9965.EPI-16-1046 Review Cancer Epidemiology, Biomarkers A Review of Prostate Cancer Genome-Wide & Prevention Association Studies (GWAS) Sarah Benafif1, Zsofia Kote-Jarai1, and Rosalind A. Eeles1,2, on behalf of the PRACTICAL Consortium Abstract Prostate cancer is the most common cancer in men in for population-based risk stratification to target prostate Europe and the United States. The genetic heritability of cancer screening to men with an increased genetic risk of prostate cancer is contributed to by both rarely occurring disease development, while for those who develop prostate genetic variants with higher penetrance and moderate cancer, identifying genetic variants could allow treatment to commonly occurring variants conferring lower risks. to be tailored based on a genetic profile in the early disease The number of identified variants belonging to the latter setting. Functional studies of identified variants are needed category has increased dramatically in the last 10 years with to fully understand underlying mechanisms of disease the development of the genome-wide association study and identify novel targets for treatment. This review will (GWAS) and the collaboration of international consortia outline the GWAS carried out in prostate cancer and the that have led to the sharing of large-scale genotyping data. common variants identified so far, and how these may be Over 40 prostate cancer GWAS have been reported, with utilized clinically in the screening for and management of approximately 170 common variants now identified. prostate cancer. Cancer Epidemiol Biomarkers Prev; 27(8); 845–57. Clinical utility of these variants could include strategies Ó2018 AACR.
    [Show full text]
  • Checkpoint-Dependent and Independent Roles of the Werner
    12628–12639 Nucleic Acids Research, 2014, Vol. 42, No. 20 Published online 28 October 2014 doi: 10.1093/nar/gku1022 Checkpoint-dependent and independent roles of the Werner syndrome protein in preserving genome integrity in response to mild replication stress Giorgia Basile1,2,†, Giuseppe Leuzzi1,2,†, Pietro Pichierri2,3 and Annapaola Franchitto1,2,* 1Section of Molecular Epidemiology, Department of Environment and Primary Prevention, Istituto Superiore di Sanita,` Viale Regina Elena, 299-00161 Rome, Italy, 2Genome Stability Group, Istituto Superiore di Sanita,` Viale Regina Elena, 299-00161 Rome, Italy and 3Section of Experimental and Computational Carcinogenesis, Department of Environment and Primary Prevention, Istituto Superiore di Sanita,` Viale Regina Elena, 299-00161 Rome, Italy Received July 22, 2014; Revised October 08, 2014; Accepted October 09, 2014 ABSTRACT agents perturbing DNA replication (1–3). Based on WRN enzymatic activities and substrate preferences in vitro,it Werner syndrome (WS) is a human chromosomal in- is thought that WRN may participate in multiple DNA stability disorder associated with cancer predispo- metabolic pathways in vivo, such as replication, recombi- sition and caused by mutations in the WRN gene. nation and repair (4–6). WRN has been implicated in the WRN helicase activity is crucial in limiting breakage correct and fruitful recovery from replication fork arrest at common fragile sites (CFS), which are the prefer- (1,7–8). Given a coordinate action of WRN and DNA poly- ential targets of genome instability in precancerous merase delta in the replication of DNA substrates contain- lesions. However, the precise function of WRN in re- ing G4 tetraplex structures (9), the crucial requirement of sponse to mild replication stress, like that commonly the WRN helicase activity in maintaining common frag- used to induce breaks at CFS, is still missing.
    [Show full text]
  • Regulation of ATR Substrate Selection by Rad17-Dependent Loading of Rad9 Complexes Onto Chromatin
    Downloaded from genesdev.cshlp.org on September 29, 2021 - Published by Cold Spring Harbor Laboratory Press Regulation of ATR substrate selection by Rad17-dependent loading of Rad9 complexes onto chromatin Lee Zou,1,2 David Cortez,1,2 and Stephen J. Elledge1–4 1Verna and Marrs McLean Department of Biochemistry and Molecular Biology, 2Howard Hughes Medical Institute, and 3Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA Cells respond to DNA damage by activating a network of signaling pathways that control cell cycle progression and DNA repair. Genetic studies in yeast suggested that several checkpoint proteins, including the RFC-related Rad17 protein, and the PCNA-related Rad1–Rad9–Hus1 protein complex might function as sensors of DNA damage. In this study, we show that the human Rad17 protein recruits the Rad9 protein complex onto chromatin after damage. Rad17 binds to chromatin prior to damage and is phosphorylated by ATR on chromatin after damage but Rad17’s phosphorylation is not required for Rad9 loading onto chromatin. The chromatin associations of Rad17 and ATR are largely independent, which suggests that they localize to DNA damage independently. Furthermore, the phosphorylation of Rad17 requires Hus1, suggesting that the Rad1–Rad9–Hus1 complex recruited by Rad17 enables ATR to recognize its substrates. Our data are consistent with a model in which multiple checkpoint protein complexes localize to sites of DNA damage independently and interact to trigger the checkpoint-signaling cascade. [Key Words: Rad17; Rad1–Rad9–Hus1 complex; ATR; Chk1; checkpoint] Received October 2, 2001; revised version accepted November 26, 2001.
    [Show full text]
  • Eukaryotic DNA Replication & Genome Maintenance
    Eukaryotic DNA Replication & Genome Maintenance Session 1 NEW APPROACHES AND PERSPECTIVES MONDAY 9/9/2013, 7:30 PM A. van Oijen / K. Labib # lname Title Talk Length 1 van Oijen Single-molecule studies of DNA replication 12 2 Remus Origin specificity during budding yeast DNA replication in vitro 12 3 Urban Mapping DNA replication origins in the human genome 12 4 Mechali DNA replication origins profiling—Genetic and epigenetic features, and relationships with cell 12 identity 5 Ni Subunits of the origin recognition complex bind BubR1 and are essential for kinetochore function 12 during mitosis in human cells 6 Labib Regulation of the replisome by ubiquitin and SUMO 12 7 Koren Random replication of the inactive X chromosome 12 8 Xie Replication-dependent DNA fragility and adaptive evolution in natural populations 12 9 Raghuraman Replication origins and the generation of palindromic amplifications in yeast 12 10 Dutta MicroDNAs and microdeletions—Genomic instability in vertebrate cells 12 Session 2 ORIGIN SELECTION AND LICENSING TUESDAY 9/10/2013, 9:00 AM M. Aladjem / S. Bell # lname Title Talk Length 11 Aladjem Regulatory interactions governing replication initiation patterns in human cells 12 12 Petryk Sequencing of Okazaki fragments allows determination of replication fork polarity and origin 12 location and efficiency in the human genome 13 Debatisse Chk1 or Rad51 deficiency perturbs replication dynamics via p53R2-mediated modulation of dNTP 12 availability 14 Liachko Dual modes of DNA replication initiation in the methylotrophic yeast
    [Show full text]
  • Hunter Et Al CLSPN
    bioRxiv preprint doi: https://doi.org/10.1101/358291; this version posted June 28, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Regulation of checkpoint kinase signalling and tumorigenesis by the NF-κB regulated gene, CLSPN Jill E. Hunter1, Jacqueline A. Butterworth1, Helene Sellier1, Saimir Luli2, Achilleas Floudas2, Adam J. Moore1, Huw D. Thomas3, Kirsteen J. Campbell4, Niall S. Kenneth1, Robson T. Chiremba1, Dina Tiniakos2, Andrew M. Knight2, Benjamin E. Gewurz5, Fiona Oakley2, Michelle D. Garrett6,7, Ian Collins7, and Neil D. Perkins1* 1Institute for Cell and Molecular Biosciences 2Institute of Cellular Medicine 3Northern Institute for Cancer Research Faculty of Medical Sciences Newcastle University Newcastle Upon Tyne, NE2 4HH, UK 4The Beatson Institute for Cancer Research Glasgow, G61 1BD, UK 5Brigham and Women's Hospital, Boston, MA 02115, USA 6School of Biosciences, University of Kent Canterbury, CT2 7NJ, UK 7The Institute of Cancer Research Sutton, SM2 5NG, UK * corresponding author Tel. 0191 2088866 Fax. 0191 2087424 Email: [email protected] Running Title: NF-κB regulation of CHK1 inhibitor sensitivity Word count: 1 bioRxiv preprint doi: https://doi.org/10.1101/358291; this version posted June 28, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract Inhibition of the tumour promoting activities of NF-κB by cell signalling pathways has been proposed as a natural mechanism to limit the development of cancer. However, there has been a lack of evidence for these effects in vivo.
    [Show full text]
  • Cdna Cloning and Gene Mapping of Human Homologs For
    GENOMICS 54, 424–436 (1998) ARTICLE NO. GE985587 cDNA Cloning and Gene Mapping of Human Homologs for Schizosaccharomyces pombe rad17, rad1, and hus1 and Cloning of Homologs from Mouse, Caenorhabditis elegans, and Drosophila melanogaster Frank B. Dean,*,1 Lubing Lian,† and Mike O’Donnell*,‡ *The Rockefeller University and ‡The Howard Hughes Medical Institute, 1230 York Avenue, New York, New York 10021; and †Myriad Genetics, Inc., 390 Wakara Way, Salt Lake City, Utah 84108 Received July 13, 1998; accepted September 17, 1998 INTRODUCTION Mutations in DNA repair/cell cycle checkpoint genes can lead to the development of cancer. The cloning of The progression of a eukaryotic cell through the human homologs of yeast DNA repair/cell cycle check- stages of the cell cycle can be arrested if the events of point genes should yield candidates for human tumor the previous stage of the cell cycle, such as DNA rep- suppressor genes as well as identifying potential tar- lication, have not been completed or if the DNA has gets for cancer therapy. The Schizosaccharomyces sustained some type of damage. The controls on cell pombe genes rad17, rad1, and hus1 have been identi- cycle progression are termed checkpoints (Hartwell fied as playing roles in DNA repair and cell cycle and Weinert, 1989), and they are able to detect checkpoint control pathways. We have cloned the whether the processes of the individual stages of the cDNA for the human homolog of S. pombe rad17, cell cycle have been completed and whether the DNA is RAD17, which localizes to chromosomal location 5q13 intact or in need of repair.
    [Show full text]
  • Somatic Mutation of the Cohesin Complex Subunit Confers Therapeutic Vulnerabilities in Cancer
    Somatic mutation of the cohesin complex subunit confers therapeutic vulnerabilities in cancer Yunhua Liu, … , Guang Ji, Xiongbin Lu J Clin Invest. 2018;128(7):2951-2965. https://doi.org/10.1172/JCI98727. Research Article Oncology Therapeutics A synthetic lethality–based strategy has been developed to identify therapeutic targets in cancer harboring tumor- suppressor gene mutations, as exemplified by the effectiveness of poly ADP-ribose polymerase (PARP) inhibitors in BRCA1/2-mutated tumors. However, many synthetic lethal interactors are less reliable due to the fact that such genes usually do not perform fundamental or indispensable functions in the cell. Here, we developed an approach to identifying the “essential lethality” arising from these mutated/deleted essential genes, which are largely tolerated in cancer cells due to genetic redundancy. We uncovered the cohesion subunit SA1 as a putative synthetic-essential target in cancers carrying inactivating mutations of its paralog, SA2. In SA2-deficient Ewing sarcoma and bladder cancer, further depletion of SA1 profoundly and specifically suppressed cancer cell proliferation, survival, and tumorigenic potential. Mechanistically, inhibition of SA1 in the SA2-mutated cells led to premature chromatid separation, dramatic extension of mitotic duration, and consequently, lethal failure of cell division. More importantly, depletion of SA1 rendered those SA2- mutated cells more susceptible to DNA damage, especially double-strand breaks (DSBs), due to reduced functionality of DNA repair. Furthermore,
    [Show full text]