POLRMT Regulates the Switch Between Replication Primer
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(PI3K(P110 )) Directly Regulates Key Components of the Z-Disc And
THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 286, NO. 35, pp. 30837–30846, September 2, 2011 © 2011 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in the U.S.A. Phosphoinositide 3-Kinase (PI3K(p110␣)) Directly Regulates Key Components of the Z-disc and Cardiac Structure*□S Received for publication, June 13, 2011, and in revised form, July 7, 2011 Published, JBC Papers in Press, July 11, 2011, DOI 10.1074/jbc.M111.271684 Ashley J. Waardenberg‡§, Bianca C. Bernardo¶, Dominic C. H. Ngʈ, Peter R. Shepherd**‡‡, Nelly Cemerlang¶, Mauro Sbroggio` §§, Christine A. Wells‡¶¶, Brian P. Dalrymple§, Mara Brancaccio§§, Ruby C. Y. Linʈʈ1, and Julie R. McMullen¶1,2 From the ‡Eskitis Institute for Cell and Molecular Therapies, Griffith University, Nathan, Queensland, 4111, Australia, §Commonwealth Scientific and Industrial Research Organisation, Food Futures Flagship, Queensland Bioscience Precinct, St. Lucia, Queensland, 4067, Australia, the ¶Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, 8008, Australia, the ʈDepartment of Biochemistry and Molecular Biology, Bio21 Institute, University of Melbourne, Melbourne, Victoria, 3010, Australia, the **Department of Molecular Medicine, University of Auckland, Grafton, Auckland, 1142, New Zealand, the ‡‡Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Grafton, Auckland, 1142, New Zealand, the §§Department of Genetics, Biology, and Biochemistry, University of Torino, Molecular Biotechnology Center, Torino, 10126, Italy, the ¶¶Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland, 4072, Australia, and the ʈʈRamaciotti Centre for Gene Function Analysis and the School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales, 2052, Australia Downloaded from Maintenance of cardiac structure and Z-disc signaling are key from PI3K transgenic mice. -
Progressive Increase in Mtdna 3243A>G Heteroplasmy Causes Abrupt
Progressive increase in mtDNA 3243A>G PNAS PLUS heteroplasmy causes abrupt transcriptional reprogramming Martin Picarda, Jiangwen Zhangb, Saege Hancockc, Olga Derbenevaa, Ryan Golhard, Pawel Golike, Sean O’Hearnf, Shawn Levyg, Prasanth Potluria, Maria Lvovaa, Antonio Davilaa, Chun Shi Lina, Juan Carlos Perinh, Eric F. Rappaporth, Hakon Hakonarsonc, Ian A. Trouncei, Vincent Procaccioj, and Douglas C. Wallacea,1 aCenter for Mitochondrial and Epigenomic Medicine, Children’s Hospital of Philadelphia and the Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104; bSchool of Biological Sciences, The University of Hong Kong, Hong Kong, People’s Republic of China; cTrovagene, San Diego, CA 92130; dCenter for Applied Genomics, Division of Genetics, Department of Pediatrics, and hNucleic Acid/Protein Research Core Facility, Children’s Hospital of Philadelphia, Philadelphia, PA 19104; eInstitute of Genetics and Biotechnology, Warsaw University, 00-927, Warsaw, Poland; fMorton Mower Central Research Laboratory, Sinai Hospital of Baltimore, Baltimore, MD 21215; gGenomics Sevices Laboratory, HudsonAlpha Institute for Biotechnology, Huntsville, AL 35806; iCentre for Eye Research Australia, Royal Victorian Eye and Ear Hospital, East Melbourne, VIC 3002, Australia; and jDepartment of Biochemistry and Genetics, National Center for Neurodegenerative and Mitochondrial Diseases, Centre Hospitalier Universitaire d’Angers, 49933 Angers, France Contributed by Douglas C. Wallace, August 1, 2014 (sent for review May -
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. -
4-6 Weeks Old Female C57BL/6 Mice Obtained from Jackson Labs Were Used for Cell Isolation
Methods Mice: 4-6 weeks old female C57BL/6 mice obtained from Jackson labs were used for cell isolation. Female Foxp3-IRES-GFP reporter mice (1), backcrossed to B6/C57 background for 10 generations, were used for the isolation of naïve CD4 and naïve CD8 cells for the RNAseq experiments. The mice were housed in pathogen-free animal facility in the La Jolla Institute for Allergy and Immunology and were used according to protocols approved by the Institutional Animal Care and use Committee. Preparation of cells: Subsets of thymocytes were isolated by cell sorting as previously described (2), after cell surface staining using CD4 (GK1.5), CD8 (53-6.7), CD3ε (145- 2C11), CD24 (M1/69) (all from Biolegend). DP cells: CD4+CD8 int/hi; CD4 SP cells: CD4CD3 hi, CD24 int/lo; CD8 SP cells: CD8 int/hi CD4 CD3 hi, CD24 int/lo (Fig S2). Peripheral subsets were isolated after pooling spleen and lymph nodes. T cells were enriched by negative isolation using Dynabeads (Dynabeads untouched mouse T cells, 11413D, Invitrogen). After surface staining for CD4 (GK1.5), CD8 (53-6.7), CD62L (MEL-14), CD25 (PC61) and CD44 (IM7), naïve CD4+CD62L hiCD25-CD44lo and naïve CD8+CD62L hiCD25-CD44lo were obtained by sorting (BD FACS Aria). Additionally, for the RNAseq experiments, CD4 and CD8 naïve cells were isolated by sorting T cells from the Foxp3- IRES-GFP mice: CD4+CD62LhiCD25–CD44lo GFP(FOXP3)– and CD8+CD62LhiCD25– CD44lo GFP(FOXP3)– (antibodies were from Biolegend). In some cases, naïve CD4 cells were cultured in vitro under Th1 or Th2 polarizing conditions (3, 4). -
Core Human Mitochondrial Transcription Apparatus Is a Regulated Two-Component System in Vitro
Core human mitochondrial transcription apparatus is a regulated two-component system in vitro Timothy E. Shutta, Maria F. Lodeirob, Justin Cotneya, Craig E. Cameronb, and Gerald S. Shadela,c,1 aDepartment of Pathology, Yale University School of Medicine, 310 Cedar Street, P.O. Box 208023, New Haven, CT 06520-8023; bDepartment of Biochemistry and Molecular Biology, Pennsylvania State University, 201 Althouse Laboratory, University Park, PA 16802; and cDepartment of Genetics, Yale University School of Medicine, 333 Cedar Street, P.O. Box 208005, New Haven, CT 06520-8005 Edited by David A. Clayton, Howard Hughes Medical Institute, Ashburn, VA, and accepted by the Editorial Board May 25, 2010 (received for review September 15, 2009) The core human mitochondrial transcription apparatus is currently ribosomal proteins to generate mitochondrial ribosomes. Finally, regarded as an obligate three-component system comprising the transcripts from the LSP are also used as primers for mtDNA bacteriophage T7-related mitochondrial RNA polymerase, the rRNA replication (5, 6), thus LSP transcription serves a dual role in methyltransferase-related transcription factor, h-mtTFB2, and the gene expression and mtDNA maintenance (7, 8). high mobility group box transcription/DNA-packaging factor, Much effort has been devoted to identifying the core machin- h-mtTFA/TFAM. Using a faithful recombinant human mitochondrial ery needed for mitochondrial transcription in humans as a critical transcription system from Escherichia coli, we demonstrate that step toward understanding the mechanism of human mitochon- specific initiation from the mtDNA promoters, LSP and HSP1, only drial gene expression and replication in vivo and its role in human requires mitochondrial RNA polymerase and h-mtTFB2 in vitro. -
Identification of Transcriptomic Differences Between Lower
International Journal of Molecular Sciences Article Identification of Transcriptomic Differences between Lower Extremities Arterial Disease, Abdominal Aortic Aneurysm and Chronic Venous Disease in Peripheral Blood Mononuclear Cells Specimens Daniel P. Zalewski 1,*,† , Karol P. Ruszel 2,†, Andrzej St˛epniewski 3, Dariusz Gałkowski 4, Jacek Bogucki 5 , Przemysław Kołodziej 6 , Jolanta Szyma ´nska 7 , Bartosz J. Płachno 8 , Tomasz Zubilewicz 9 , Marcin Feldo 9,‡ , Janusz Kocki 2,‡ and Anna Bogucka-Kocka 1,‡ 1 Chair and Department of Biology and Genetics, Medical University of Lublin, 4a Chod´zkiSt., 20-093 Lublin, Poland; [email protected] 2 Chair of Medical Genetics, Department of Clinical Genetics, Medical University of Lublin, 11 Radziwiłłowska St., 20-080 Lublin, Poland; [email protected] (K.P.R.); [email protected] (J.K.) 3 Ecotech Complex Analytical and Programme Centre for Advanced Environmentally Friendly Technologies, University of Marie Curie-Skłodowska, 39 Gł˛ebokaSt., 20-612 Lublin, Poland; [email protected] 4 Department of Pathology and Laboratory Medicine, Rutgers-Robert Wood Johnson Medical School, One Robert Wood Johnson Place, New Brunswick, NJ 08903-0019, USA; [email protected] 5 Chair and Department of Organic Chemistry, Medical University of Lublin, 4a Chod´zkiSt., Citation: Zalewski, D.P.; Ruszel, K.P.; 20-093 Lublin, Poland; [email protected] St˛epniewski,A.; Gałkowski, D.; 6 Laboratory of Diagnostic Parasitology, Chair and Department of Biology and Genetics, Medical University of Bogucki, J.; Kołodziej, P.; Szyma´nska, Lublin, 4a Chod´zkiSt., 20-093 Lublin, Poland; [email protected] J.; Płachno, B.J.; Zubilewicz, T.; Feldo, 7 Department of Integrated Paediatric Dentistry, Chair of Integrated Dentistry, Medical University of Lublin, M.; et al. -
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 -
Intimate Relations—Mitochondria and Ageing
International Journal of Molecular Sciences Review Intimate Relations—Mitochondria and Ageing Michael Webb and Dionisia P. Sideris * Mitobridge Inc., an Astellas Company, 1030 Massachusetts Ave, Cambridge, MA 02138, USA; [email protected] * Correspondence: [email protected] Received: 29 August 2020; Accepted: 6 October 2020; Published: 14 October 2020 Abstract: Mitochondrial dysfunction is associated with ageing, but the detailed causal relationship between the two is still unclear. Wereview the major phenomenological manifestations of mitochondrial age-related dysfunction including biochemical, regulatory and energetic features. We conclude that the complexity of these processes and their inter-relationships are still not fully understood and at this point it seems unlikely that a single linear cause and effect relationship between any specific aspect of mitochondrial biology and ageing can be established in either direction. Keywords: mitochondria; ageing; energetics; ROS; gene regulation 1. Introduction The last two decades have witnessed a dramatic transformation in our view of mitochondria, their basic biology and functions. While still regarded as functioning primarily as the eukaryotic cell’s generator of energy in the form of adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide (reduced form; NADH), mitochondria are now recognized as having a plethora of functions, including control of apoptosis, regulation of calcium, forming a signaling hub and the synthesis of various bioactive molecules. Their biochemical functions beyond ATP supply include biosynthesis of lipids and amino acids, formation of iron sulphur complexes and some stages of haem biosynthesis and the urea cycle. They exist as a dynamic network of organelles that under normal circumstances undergo a constant series of fission and fusion events in which structural, functional and encoding (mtDNA) elements are subject to redistribution throughout the network. -
Common Chemical Inductors of Replication Stress: Focus on Cell-Based Studies
biomolecules Review Common Chemical Inductors of Replication Stress: Focus on Cell-Based Studies Eva Vesela 1,2, Katarina Chroma 1, Zsofia Turi 1 and Martin Mistrik 1,* 1 Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Hnevotinska 5, Olomouc 779 00, Czech Republic; [email protected] (E.V.); [email protected] (K.C.); zsofi[email protected] (Z.T.) 2 MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK * Correspondence: [email protected]; Tel.: +420-585-634-170 Academic Editor: Rob de Bruin Received: 25 November 2016; Accepted: 10 February 2017; Published: 21 February 2017 Abstract: DNA replication is a highly demanding process regarding the energy and material supply and must be precisely regulated, involving multiple cellular feedbacks. The slowing down or stalling of DNA synthesis and/or replication forks is referred to as replication stress (RS). Owing to the complexity and requirements of replication, a plethora of factors may interfere and challenge the genome stability, cell survival or affect the whole organism. This review outlines chemical compounds that are known inducers of RS and commonly used in laboratory research. These compounds act on replication by direct interaction with DNA causing DNA crosslinks and bulky lesions (cisplatin), chemical interference with the metabolism of deoxyribonucleotide triphosphates (hydroxyurea), direct inhibition of the activity of replicative DNA polymerases (aphidicolin) and interference with enzymes dealing with topological DNA stress (camptothecin, etoposide). As a variety of mechanisms can induce RS, the responses of mammalian cells also vary. Here, we review the activity and mechanism of action of these compounds based on recent knowledge, accompanied by examples of induced phenotypes, cellular readouts and commonly used doses. -
Human Mitochondrial RNA Polymerase Primes Lagging-Strand DNA Synthesis in Vitro
Human mitochondrial RNA polymerase primes lagging-strand DNA synthesis in vitro Sjoerd Wanrooij*†, Javier Miralles Fuste´ *, Ge´ raldine Farge*, Yonghong Shi*†, Claes M. Gustafsson*†‡, and Maria Falkenberg*†‡ *Division of Metabolic Diseases, Karolinska Institutet, Novum, SE-141 86 Stockholm, Sweden; and †Department of Medical Biochemistry and Cell Biology, Go¨teborg University, Box 440, SE-405 30 Go¨teborg, Sweden Communicated by I. Robert Lehman, Stanford University School of Medicine, Stanford, CA, June 4, 2008 (received for review March 14, 2008) The mitochondrial transcription machinery synthesizes the RNA strand promoters (LSP) generates transcripts of almost genomic primers required for initiation of leading-strand DNA synthesis in length, which are processed to yield the individual mRNA and mammalian mitochondria. RNA primers are also required for ini- tRNA molecules (10, 11). LSP-dependent transcription also tiation of lagging-strand DNA synthesis, but the responsible en- generates the primers needed at OH (12), and RNA attached to zyme has so far remained elusive. Here, we present a series of the newly synthesized H strand has been detected in both mouse observations that suggests that mitochondrial RNA polymerase and human cells (13). Primer formation may include an RNase (POLRMT) can act as lagging-strand primase in mammalian cells. MRP-dependent cleavage of the transcribed L-strand RNA at POLRMT is highly processive on double-stranded DNA, but synthe- the initiation sites of DNA replication (14). Alternatively, spe- sizes RNA primers with a length of 25 to 75 nt on a single-stranded cific mtDNA sequences may direct premature transcription template. The short RNA primers synthesized by POLRMT are used termination and primer formation near OH (15). -
Supplementary Data.Xlsx
Electronic Supplementary Material (ESI) for Molecular BioSystems. This journal is © The Royal Society of Chemistry 2016 Average Average spectral spectral Fold UniProt IDGene Protein Name counts- counts- enrichm negative positive ent sample sample P12821 ACE HUMAN - ACE Angiotensin-converting enzyme 0 79.75 #DIV/0! Q71U36 TBA1A HUMAN - TUBA1A Tubulin alpha-1A chain 0 59.5 #DIV/0! P17812 PYRG1 HUMAN - CTPS1 CTP synthase 1 0 43.5 #DIV/0! P23921 RIR1 HUMAN - RRM1 Ribonucleoside-diphosphate reductase large subunit 0 35 #DIV/0! P49915GUAA HUMAN - GMPS GMP synthase 0 30.5 #DIV/0! P30153 2AAA HUMAN - PPP2R1A Serine/threonine-protein phosphatase 2A 65 kDa0 regulatory subunit29 A#DIV/0! alpha isoform P55786 PSA HUMAN - NPEPPS Puromycin-sensitive aminopeptidase 0 28.75 #DIV/0! O43143 DHX15 HUMAN - DHX15 Putative pre-mRNA-splicing factor ATP-dependent RNA0 helicase28.25 DHX15#DIV/0! P15170 ERF3A HUMAN - GSPT1 Eukaryotic peptide chain release factor GTP-binding0 subunit ERF3A24.75 #DIV/0! P09874PARP1HUMAN - PARP1 Poly 0 23.5 #DIV/0! Q9BXJ9 NAA15 HUMAN - NAA15 N-alpha-acetyltransferase 15, NatA auxiliary subunit0 23 #DIV/0! B0V043 B0V043 HUMAN - VARS Valyl-tRNA synthetase 0 20 #DIV/0! Q86VP6 CAND1 HUMAN - CAND1 Cullin-associated NEDD8-dissociated protein 1 0 19.5 #DIV/0! P04080CYTB HUMAN - CSTB Cystatin-B 0 19 #DIV/0! Q93009 UBP7 HUMAN - USP7 Ubiquitin carboxyl-terminal hydrolase 7 0 18 #DIV/0! Q9Y2L1 RRP44 HUMAN - DIS3 Exosome complex exonuclease RRP44 0 18 #DIV/0! Q13748 TBA3C HUMAN - TUBA3D Tubulin alpha-3C/D chain 0 18 #DIV/0! P29144 TPP2 HUMAN -
A Novel DNA Primase-Helicase Pair Encoded by Sccmec Elements Aleksandra Bebel†, Melissa a Walsh, Ignacio Mir-Sanchis‡, Phoebe a Rice*
RESEARCH ARTICLE A novel DNA primase-helicase pair encoded by SCCmec elements Aleksandra Bebel†, Melissa A Walsh, Ignacio Mir-Sanchis‡, Phoebe A Rice* Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, United States Abstract Mobile genetic elements (MGEs) are a rich source of new enzymes, and conversely, understanding the activities of MGE-encoded proteins can elucidate MGE function. Here, we biochemically characterize three proteins encoded by a conserved operon carried by the Staphylococcal Cassette Chromosome (SCCmec), an MGE that confers methicillin resistance to Staphylococcus aureus, creating MRSA strains. The first of these proteins, CCPol, is an active A-family DNA polymerase. The middle protein, MP, binds tightly to CCPol and confers upon it the ability to synthesize DNA primers de novo. The CCPol-MP complex is therefore a unique primase- polymerase enzyme unrelated to either known primase family. The third protein, Cch2, is a 3’-to-5’ helicase. Cch2 additionally binds specifically to a dsDNA sequence downstream of its gene that is also a preferred initiation site for priming by CCPol-MP. Taken together, our results suggest that this is a functional replication module for SCCmec. *For correspondence: Introduction [email protected] Staphylococcus aureus is a dangerous human pathogen, due in part to the emergence of multi- drug-resistant strains such as MRSA (methicillin-resistant S. aureus). MRSA strains have acquired † Present address: Phage resistance to b-lactam antibiotics (including methicillin) mainly through horizontal gene transfer of a Consultants, Gdynia, Poland; mobile genomic island called staphylococcal cassette chromosome (SCC) (Moellering, 2012). ‡Umea˚ University, Umea˚ , SCCmec is a variant of SCC that carries a methicillin resistance gene, mecA.