DNA Double-Strand Break Formation and Signalling in Response to Transcription-Blocking Topoisomerase I Complexes Agnese Cristini
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Large XPF-Dependent Deletions Following Misrepair of a DNA Double Strand Break Are Prevented by the RNA:DNA Helicase Senataxin
www.nature.com/scientificreports OPEN Large XPF-dependent deletions following misrepair of a DNA double strand break are prevented Received: 26 October 2017 Accepted: 9 February 2018 by the RNA:DNA helicase Published: xx xx xxxx Senataxin Julien Brustel1, Zuzanna Kozik1, Natalia Gromak2, Velibor Savic3,4 & Steve M. M. Sweet1,5 Deletions and chromosome re-arrangements are common features of cancer cells. We have established a new two-component system reporting on epigenetic silencing or deletion of an actively transcribed gene adjacent to a double-strand break (DSB). Unexpectedly, we fnd that a targeted DSB results in a minority (<10%) misrepair event of kilobase deletions encompassing the DSB site and transcribed gene. Deletions are reduced upon RNaseH1 over-expression and increased after knockdown of the DNA:RNA helicase Senataxin, implicating a role for DNA:RNA hybrids. We further demonstrate that the majority of these large deletions are dependent on the 3′ fap endonuclease XPF. DNA:RNA hybrids were detected by DNA:RNA immunoprecipitation in our system after DSB generation. These hybrids were reduced by RNaseH1 over-expression and increased by Senataxin knock-down, consistent with a role in deletions. Overall, these data are consistent with DNA:RNA hybrid generation at the site of a DSB, mis-processing of which results in genome instability in the form of large deletions. DNA is the target of numerous genotoxic attacks that result in diferent types of damage. DNA double-strand breaks (DSBs) occur at low frequency, compared with single-strand breaks and other forms of DNA damage1, however DSBs pose the risk of translocations and deletions and their repair is therefore essential to cell integrity. -
A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated. -
Supp Material.Pdf
Simon et al. Supplementary information: Table of contents p.1 Supplementary material and methods p.2-4 • PoIy(I)-poly(C) Treatment • Flow Cytometry and Immunohistochemistry • Western Blotting • Quantitative RT-PCR • Fluorescence In Situ Hybridization • RNA-Seq • Exome capture • Sequencing Supplementary Figures and Tables Suppl. items Description pages Figure 1 Inactivation of Ezh2 affects normal thymocyte development 5 Figure 2 Ezh2 mouse leukemias express cell surface T cell receptor 6 Figure 3 Expression of EZH2 and Hox genes in T-ALL 7 Figure 4 Additional mutation et deletion of chromatin modifiers in T-ALL 8 Figure 5 PRC2 expression and activity in human lymphoproliferative disease 9 Figure 6 PRC2 regulatory network (String analysis) 10 Table 1 Primers and probes for detection of PRC2 genes 11 Table 2 Patient and T-ALL characteristics 12 Table 3 Statistics of RNA and DNA sequencing 13 Table 4 Mutations found in human T-ALLs (see Fig. 3D and Suppl. Fig. 4) 14 Table 5 SNP populations in analyzed human T-ALL samples 15 Table 6 List of altered genes in T-ALL for DAVID analysis 20 Table 7 List of David functional clusters 31 Table 8 List of acquired SNP tested in normal non leukemic DNA 32 1 Simon et al. Supplementary Material and Methods PoIy(I)-poly(C) Treatment. pIpC (GE Healthcare Lifesciences) was dissolved in endotoxin-free D-PBS (Gibco) at a concentration of 2 mg/ml. Mice received four consecutive injections of 150 μg pIpC every other day. The day of the last pIpC injection was designated as day 0 of experiment. -
Supplementary Table S4. FGA Co-Expressed Gene List in LUAD
Supplementary Table S4. FGA co-expressed gene list in LUAD tumors Symbol R Locus Description FGG 0.919 4q28 fibrinogen gamma chain FGL1 0.635 8p22 fibrinogen-like 1 SLC7A2 0.536 8p22 solute carrier family 7 (cationic amino acid transporter, y+ system), member 2 DUSP4 0.521 8p12-p11 dual specificity phosphatase 4 HAL 0.51 12q22-q24.1histidine ammonia-lyase PDE4D 0.499 5q12 phosphodiesterase 4D, cAMP-specific FURIN 0.497 15q26.1 furin (paired basic amino acid cleaving enzyme) CPS1 0.49 2q35 carbamoyl-phosphate synthase 1, mitochondrial TESC 0.478 12q24.22 tescalcin INHA 0.465 2q35 inhibin, alpha S100P 0.461 4p16 S100 calcium binding protein P VPS37A 0.447 8p22 vacuolar protein sorting 37 homolog A (S. cerevisiae) SLC16A14 0.447 2q36.3 solute carrier family 16, member 14 PPARGC1A 0.443 4p15.1 peroxisome proliferator-activated receptor gamma, coactivator 1 alpha SIK1 0.435 21q22.3 salt-inducible kinase 1 IRS2 0.434 13q34 insulin receptor substrate 2 RND1 0.433 12q12 Rho family GTPase 1 HGD 0.433 3q13.33 homogentisate 1,2-dioxygenase PTP4A1 0.432 6q12 protein tyrosine phosphatase type IVA, member 1 C8orf4 0.428 8p11.2 chromosome 8 open reading frame 4 DDC 0.427 7p12.2 dopa decarboxylase (aromatic L-amino acid decarboxylase) TACC2 0.427 10q26 transforming, acidic coiled-coil containing protein 2 MUC13 0.422 3q21.2 mucin 13, cell surface associated C5 0.412 9q33-q34 complement component 5 NR4A2 0.412 2q22-q23 nuclear receptor subfamily 4, group A, member 2 EYS 0.411 6q12 eyes shut homolog (Drosophila) GPX2 0.406 14q24.1 glutathione peroxidase -
The Influence of Cell Cycle Regulation on Chemotherapy
International Journal of Molecular Sciences Review The Influence of Cell Cycle Regulation on Chemotherapy Ying Sun 1, Yang Liu 1, Xiaoli Ma 2 and Hao Hu 1,* 1 Institute of Biomedical Materials and Engineering, College of Materials Science and Engineering, Qingdao University, Qingdao 266071, China; [email protected] (Y.S.); [email protected] (Y.L.) 2 Qingdao Institute of Measurement Technology, Qingdao 266000, China; [email protected] * Correspondence: [email protected] Abstract: Cell cycle regulation is orchestrated by a complex network of interactions between proteins, enzymes, cytokines, and cell cycle signaling pathways, and is vital for cell proliferation, growth, and repair. The occurrence, development, and metastasis of tumors are closely related to the cell cycle. Cell cycle regulation can be synergistic with chemotherapy in two aspects: inhibition or promotion. The sensitivity of tumor cells to chemotherapeutic drugs can be improved with the cooperation of cell cycle regulation strategies. This review presented the mechanism of the commonly used chemotherapeutic drugs and the effect of the cell cycle on tumorigenesis and development, and the interaction between chemotherapy and cell cycle regulation in cancer treatment was briefly introduced. The current collaborative strategies of chemotherapy and cell cycle regulation are discussed in detail. Finally, we outline the challenges and perspectives about the improvement of combination strategies for cancer therapy. Keywords: chemotherapy; cell cycle regulation; drug delivery systems; combination chemotherapy; cancer therapy Citation: Sun, Y.; Liu, Y.; Ma, X.; Hu, H. The Influence of Cell Cycle Regulation on Chemotherapy. Int. J. 1. Introduction Mol. Sci. 2021, 22, 6923. https:// Chemotherapy is currently one of the main methods of tumor treatment [1]. -
The Microbiota-Produced N-Formyl Peptide Fmlf Promotes Obesity-Induced Glucose
Page 1 of 230 Diabetes Title: The microbiota-produced N-formyl peptide fMLF promotes obesity-induced glucose intolerance Joshua Wollam1, Matthew Riopel1, Yong-Jiang Xu1,2, Andrew M. F. Johnson1, Jachelle M. Ofrecio1, Wei Ying1, Dalila El Ouarrat1, Luisa S. Chan3, Andrew W. Han3, Nadir A. Mahmood3, Caitlin N. Ryan3, Yun Sok Lee1, Jeramie D. Watrous1,2, Mahendra D. Chordia4, Dongfeng Pan4, Mohit Jain1,2, Jerrold M. Olefsky1 * Affiliations: 1 Division of Endocrinology & Metabolism, Department of Medicine, University of California, San Diego, La Jolla, California, USA. 2 Department of Pharmacology, University of California, San Diego, La Jolla, California, USA. 3 Second Genome, Inc., South San Francisco, California, USA. 4 Department of Radiology and Medical Imaging, University of Virginia, Charlottesville, VA, USA. * Correspondence to: 858-534-2230, [email protected] Word Count: 4749 Figures: 6 Supplemental Figures: 11 Supplemental Tables: 5 1 Diabetes Publish Ahead of Print, published online April 22, 2019 Diabetes Page 2 of 230 ABSTRACT The composition of the gastrointestinal (GI) microbiota and associated metabolites changes dramatically with diet and the development of obesity. Although many correlations have been described, specific mechanistic links between these changes and glucose homeostasis remain to be defined. Here we show that blood and intestinal levels of the microbiota-produced N-formyl peptide, formyl-methionyl-leucyl-phenylalanine (fMLF), are elevated in high fat diet (HFD)- induced obese mice. Genetic or pharmacological inhibition of the N-formyl peptide receptor Fpr1 leads to increased insulin levels and improved glucose tolerance, dependent upon glucagon- like peptide-1 (GLP-1). Obese Fpr1-knockout (Fpr1-KO) mice also display an altered microbiome, exemplifying the dynamic relationship between host metabolism and microbiota. -
DNA Topoisomerase I and DNA Gyrase As Targets for TB Therapy, Drug Discov Today (2016), J.Drudis.2016.11.006
Drug Discovery Today Volume 00, Number 00 November 2016 REVIEWS GENE TO SCREEN DNA topoisomerase I and DNA gyrase as targets for TB therapy Reviews 1,2 1 3 Valakunja Nagaraja , Adwait A. Godbole , Sara R. Henderson and 3 Anthony Maxwell 1 Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560 012, India 2 Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore 560064, India 3 Department of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK Tuberculosis (TB) is the deadliest bacterial disease in the world. New therapeutic agents are urgently needed to replace existing drugs for which resistance is a significant problem. DNA topoisomerases are well-validated targets for antimicrobial and anticancer chemotherapies. Although bacterial topoisomerase I has yet to be exploited as a target for clinical antibiotics, DNA gyrase has been extensively targeted, including the highly clinically successful fluoroquinolones, which have been utilized in TB therapy. Here, we review the exploitation of topoisomerases as antibacterial targets and summarize progress in developing new agents to target DNA topoisomerase I and DNA gyrase from Mycobacterium tuberculosis. Introduction provides for a certain degree of overlap in their functions. For Although the information content of DNA is essentially indepen- instance, in Escherichia coli, there are four topoisomerases: two dent of how the DNA is knotted or twisted, the access to this type I [topoisomerase (topo) I and topo III] and two type II (DNA information depends on the topology of the DNA. DNA topoi- gyrase and topo IV). In vitro, all four enzymes are capable of DNA somerases are ubiquitous enzymes that maintain the topological relaxation, whereas, in vivo, their roles tend to be more special- homeostasis within the cell during these DNA transaction pro- ized, for example, gyrase introduces negative supercoils, whereas cesses [1–3]. -
High Expression of TOP2A Gene Predicted Poor Prognosis of Hepatocellular Carcinoma After Radical Hepatectomy
992 Original Article High expression of TOP2A gene predicted poor prognosis of hepatocellular carcinoma after radical hepatectomy Hongyu Cai1,2, Xinhua Zhu3, Fei Qian4, Bingfeng Shao2, Yuan Zhou2, Yixin Zhang2, Zhong Chen1,4 1Department of General Surgery, The First Affiliated Hospital of Soochow University, Soochow 215006, China; 2Department of Hepatobiliary Surgery, Nantong Tumor Hospital, Nantong 226361, China; 3Department of Pathological, Affiliated Tumor Hospital of Nantong University, Nantong 226006, China; 4Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University, Nantong 226001, China Contributions: (I) Conception and design: Z Chen, H Cai; (II) Administrative support: Y Zhang, Y Zhou; (III) Provision of study materials or patients: B Shao; (IV) Collection and assembly of data: H Cai, X Zhu, F Qian; (V) Data analysis and interpretation: H Cai, X Zhu; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. Correspondence to: Zhong Chen. Department of General Surgery, The First Affiliated Hospital of Soochow University, Soochow 215006, China. Email: [email protected]. Background: Topoisomerase (DNA) II alpha (TOP2A) is the up-regulated gene of the chromosome aggregation pathway. This gene encodes DNA topoisomerase, which can control and change the topological state of DNA during transcription and replication, participate in chromosome agglutination, separation, and relieve stress from kinking. Abnormally high expression of TOP2A is often associated with active cell proliferation. In studies of breast cancer, endometrial cancer, and adrenal cancer, it was found that high expression of TOP2A suggested a poor prognosis. Methods: A total of 15 pairs of fresh hepatocellular carcinoma (HCC) specimens and adjacent tissues were assembled, and the difference of TOP2A mRNA and protein expression level between tumor and adjacent tissues was detected by RT-PCR and Western blotting analysis, respectively. -
Viable Rnaseh1 Knockout Mice Show Rnaseh1 Is Essential for R Loop Processing, Mitochondrial and Liver Function Walt F
Published online 29 April 2016 Nucleic Acids Research, 2016, Vol. 44, No. 11 5299–5312 doi: 10.1093/nar/gkw350 Viable RNaseH1 knockout mice show RNaseH1 is essential for R loop processing, mitochondrial and liver function Walt F. Lima1, Heather M. Murray1, Sagar S. Damle2, Christopher E. Hart2, Gene Hung3, Cheryl Li De Hoyos1, Xue-Hai Liang1 and Stanley T. Crooke1,* 1Department of Core Antisense Research, Ionis Pharmaceuticals Inc., 2855 Gazelle Court, Carlsbad, CA 92010, USA, 2Department of Functional Genomics, Ionis Pharmaceuticals Inc., 2855 Gazelle Court, Carlsbad, CA 92010, USA and 3Department of Antisense Drug Discovery, Ionis Pharmaceuticals Inc., 2855 Gazelle Court, Carlsbad, CA 92010, USA Received February 18, 2016; Revised March 17, 2016; Accepted April 19, 2016 ABSTRACT tant for mitochondrial DNA replication (12,13). Loss of RNase H1 function in humans results in adult-onset mito- Viable constitutive and tamoxifen inducible liver- chondrial encephalomyopathy with chronic progressive ex- specific RNase H1 knockout mice that expressed no ternal ophthalmoplegia, and multiple muscle weakness con- RNase H1 activity in hepatocytes showed increased ditions (13). R-loop levels and reduced mitochondrial encoded Human RNase H1 has a hybrid binding domain homolo- DNA and mRNA levels, suggesting impaired mito- gous to yeast RNase H1 that is separated from the catalytic chondrial R-loop processing, transcription and mito- domain by a 62-amino acid spacer region (14–16). The cat- chondrial DNA replication. These changes resulted alytic domain is highly conserved from bacteria to humans, in mitochondrial dysfunction with marked changes in and contains the key catalytic residues, as well as lysine ␣ mitochondrial fusion, fission, morphology and tran- residues within the highly basic -helical substrate binding scriptional changes reflective of mitochondrial dam- region found in E. -
Ribonuclease H1-Dependent Hepatotoxicity Caused by Locked
www.nature.com/scientificreports OPEN Ribonuclease H1-dependent hepatotoxicity caused by locked nucleic acid-modified gapmer Received: 29 March 2016 Accepted: 30 June 2016 antisense oligonucleotides Published: 27 July 2016 Takeshi Kasuya1, Shin-ichiro Hori1, Ayahisa Watanabe2, Mado Nakajima1, Yoshinari Gahara1, Masatomo Rokushima1, Toru Yanagimoto1,* & Akira Kugimiya1,* Gapmer antisense oligonucleotides cleave target RNA effectivelyin vivo, and is considered as promising therapeutics. Especially, gapmers modified with locked nucleic acid (LNA) shows potent knockdown activity; however, they also cause hepatotoxic side effects. For developing safe and effective gapmer drugs, a deeper understanding of the mechanisms of hepatotoxicity is required. Here, we investigated the cause of hepatotoxicity derived from LNA-modified gapmers. Chemical modification of gapmer’s gap region completely suppressed both knockdown activity and hepatotoxicity, indicating that the root cause of hepatotoxicity is related to intracellular gapmer activity. Gene silencing of hepatic ribonuclease H1 (RNaseH1), which catalyses gapmer-mediated RNA knockdown, strongly supressed hepatotoxic effects. Small interfering RNA (siRNA)-mediated knockdown of a target mRNA did not result in any hepatotoxic effects, while the gapmer targeting the same position on mRNA as does the siRNA showed acute toxicity. Microarray analysis revealed that several pre-mRNAs containing a sequence similar to the gapmer target were also knocked down. These results suggest that hepatotoxicity of LNA gapmer is caused by RNAseH1 activity, presumably because of off-target cleavage of RNAs inside nuclei. The 5′ and 3′ end-modified antisense oligonucleotide (ASO), gapmer, is promising therapeutic agent which can modulate target RNA expression1,2. Various types of chemical modification of ASOs have been examined to enhance nuclease resistance, to improve stability of ASO–RNA hybrids, and to increase RNA-manipulating effects. -
Product Datasheet Rnase H1 Antibody (5D10
Product Datasheet RNase H1 Antibody (5D10) H00246243-M01 Unit Size: 0.1 mg Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles. Publications: 4 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: www.novusbio.com/H00246243-M01 Updated 6/9/2020 v.20.1 Earn rewards for product reviews and publications. Submit a publication at www.novusbio.com/publications Submit a review at www.novusbio.com/reviews/destination/H00246243-M01 Page 1 of 4 v.20.1 Updated 6/9/2020 H00246243-M01 RNase H1 Antibody (5D10) Product Information Unit Size 0.1 mg Concentration Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services. Storage Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles. Clonality Monoclonal Clone 5D10 Preservative No Preservative Isotype IgG2a Kappa Purity IgG purified Buffer In 1x PBS, pH 7.4 Product Description Host Mouse Gene ID 246243 Gene Symbol RNASEH1 Species Human, Rat Specificity/Sensitivity RNASEH1 - ribonuclease H1 Immunogen RNASEH1 (NP_002927, 189 a.a. ~ 286 a.a) partial recombinant protein with GST tag. MW of the GST tag alone is 26 KDa. AACKAIEQAKTQNINKLVLYTDSMFTINGITNWVQGWKKNGWKTSAGKEVINKE DFVALERLTQGMDIQWMHVPGHSGFIGNEEADRLAREGAKQSED Notes This product is produced by and distributed for Abnova, a company based in Taiwan. Product Application Details Applications Western Blot, ELISA Recommended Dilutions Western Blot 1:500, ELISA 1:100-1:2000 Application Notes Antibody reactive against cell lysate and recombinant protein for western blot. It has also been used for ELISA. Page 2 of 4 v.20.1 Updated 6/9/2020 Images Western Blot: RNase H1 Antibody (5D10) [H00246243-M01] - RNASEH1 monoclonal antibody (M01), clone 5D10. -
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RSC Advances View Article Online PAPER View Journal | View Issue Divergent synthesis of a thiolate-based a- hydroxytropolone library with a dynamic Cite this: RSC Adv.,2019,9, 34227 bioactivity profile† Nana B. Agyemang, ab Cassandra R. Kukla,c Tiffany C. Edwards,c Qilan Li,c Madison K. Langen,d Alexandra Schaal,d Abaigeal D. Franson,c Andreu Gazquez Casals,c Katherine A. Donald,c Alice J. Yu,c Maureen J. Donlin, d Lynda A. Morrison, ce John E. Tavis c and Ryan P. Murelli *ab Here we describe a rapid and divergent synthetic route toward structurally novel aHTs functionalized with Received 15th August 2019 either one or two thioether or sulfonyl appendages. Evaluation of this library against hepatitis B and herpes Accepted 7th October 2019 simplex virus, as well as the pathogenic fungus Cryptococcus neoformans, revealed complementary DOI: 10.1039/c9ra06383h biological profiles and new lead compounds with sub-micromolar activities against each pathogen and rsc.li/rsc-advances high selectivities when compared to mammalian cell lines. Creative Commons Attribution-NonCommercial 3.0 Unported Licence. Introduction functionalities at position 4 (inset, Scheme 1). However, installing functionality at the 3 position, adjacent to the metal- a-Hydroxytropolone (aHT, 1, Fig. 1A) is a highly oxygenated binding oxygens, is more laborious using this strategy,8 and it is troponoid with 3 contiguous oxygen atoms that give it the not clear at present how the method could install functionality capacity to coordinate two metals in the active sites of many at