DOCSLIB.ORG

Characterization, Expression and Chromosomal Localization of a Human Gene Homologous to the Mouse Lsc Oncogene, with Strongest Expression in Hematopoetic Tissues

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Characterization, Expression and Chromosomal Localization of a Human Gene Homologous to the Mouse Lsc Oncogene, with Strongest Expression in Hematopoetic Tissues Oncogene (1997) 14, 1747 ± 1752 1997 Stockton Press All rights reserved 0950 ± 9232/97 $12.00 SHORT REPORT Characterization, expression and chromosomal localization of a human gene homologous to the mouse Lsc oncogene, with strongest expression in hematopoetic tissues Hans-Christian Aasheim1, Florence Pedeutour2 and Erlend B Smeland1 1Department of Immunology, Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, N-0310, Oslo, Norway; 2URA CNRS 1462, Faculty de Medecine, Avenue de Valombrose, Nice, France A human cDNA clone, denoted sub1.5, was isolated from McCormick, 1993; Quilliam et al., 1995; Denhardt, cDNA library generated from human T cells. The sub1.5 1996). The Ras family itself is involved in triggering cDNA sequence was novel and was not identical to any cell proliferation in response to mitogens and growth known cDNA sequences in the GenBank. Recently, factors via the Raf/MEK/mitogen-activated protein however, a mouse cDNA (Lsc) with high homology to kinase signalling pathway leading to a phosphoryla- sub1.5 was identi®ed, indicating that the sub1.5 sequence tion cascade which activates transcription factors to may represent the human homologue of the mouse Lsc induce gene expression (Denhardt, 1996). The Rho/Rac gene. The sub1.5 cDNA includes an open reading frame family is involved in cytoskeletal organisation and of 875 amino acids, predicting a protein with molecular focal contacs (Ridley and Hall, 1992; Ridley et al., weight of 97 kDa. Like Lsc, sub1.5 shows homology to 1992) and has been suggested to participate in growth the previous described oncogene Lbc, in particular to two factor signalling (Nobes and Hall, 1995; Coso et al., functional domains in the Lbc protein; the Dbl proto- 1995; Minden et al., 1995; Hill et al., 1995) and to be oncogene homology domain and the pleckstrin homology involved in regulation of receptor mediated endocytosis domain. Lsc is proposed to be an oncogene and is a through coated pits (Lamaze et al., 1996). member of a growing family of proteins that may The Ras superfamily of GTP-binding proteins function as activators of the Rho family GTPases. alternate between alternate active GTP-bound and Members of the Rho family regulates the polymerization inactive GDP-bound states (for review see Quilliam et of actin to produce stress ®bers. Activation of Rho al., 1995). The GTP-binding/GTPase cycles are tightly GTPases by sub1.5 is also indicated by our studies, as controlled, with guanidine nucleotide exchange factors stress ®ber formation is observed in serum-starved stable (GEFs) catalyzing their conversion to the GTP-bound NIH3T3 sub1.5 transfectants. Sub1.5 cDNA hybridizes active state and GTPase-activating proteins (GAPs) to two major transcripts of 3.5 and 5 kb size and the ensuring their return to an inactive, basal state through strongest expression is seen in hematopoietic tissues like the stimulation of GTP hydrolysis (Boguski and thymus, lymph nodes, peripheral blood leukocytes and McCormick, 1993). GEFs have been described for spleen. We also show that both puri®ed B and T cells virtually all the Ras GTPase families (Boguski and express sub1.5. In addition, our data indicate that sub1.5 McCormick, 1993). Recently, the product of the Dbl- mRNA is abundantly expressed in CD34+ human oncogene was shown to serve as a GEF for speci®c progenitor cells. Fluorescent in situ hybridisation, using members of the Rho branch of the Ras superfamily sub1.5 cDNA as a probe on human metaphases, shows (RhoA, Rac1, CDC42HS; Hart et al., 1994). It was that the sub1.5 gene is localized to chromosome shown that both the GEF activity and the transform- 19q13.13. ing activity of the Dbl protein was localized to a 240 amino acid motif between residues 498 and 738 in the Keywords: Lsc homologue; Rho GEF; Dbl-family; protein (Hart et al., 1994). An emerging group of hematopoetic expression proteins conferring homology to this 240 amino acid motif in the Dbl oncogene has been identi®ed including Vav (Katzav et al., 1989), Ect-2 (Miki et al., 1993), Lbc Members of the Ras superfamily of proteins function (Toksoz and Williams, 1994), Lfc (Whitehead et al., as molecular switches in a diversity of cellular 1995), Tim (Chan et al., 1994), Tiam (Habets et al., signalling pathways, in¯uencing processes such as 1994), Cdc24 (from Saccharomysis cerevisia; Drubin, cytoskeletal organization, development, vesicular trans- 1991), the breakpoint cluster region protein (Bcr; port, cell polarity and cell motility (Adams et al., 1990; Hariharan and Adams, 1987), and the mammalian Boguski and McCormic, 1993; Ridley et al., 1992; ras guanidine releasing factor (Shou et al., 1992). Ridley and Hall, 1992). The Ras superfamily consists All Dbl family members possess a second shared of more than 50 members that share the ability to bind domain, designated the plekcstrin homology (PH) and hydrolyze GTP. The superfamily can be divided domain. The PH domain was initially identi®ed as a into several subfamilies, designated Ras, Rho/Rac, region of sequence homology, of approximately 120 Rab, Ran, and Arf/Sar (For review see Boguski and amino acids, which is duplicated in pleckstrin (reviewed in Lemmon et al., 1996; Gibson et al., 1994). Pleckstrin is the major substrate of protein kinase C in platelets. Correspondence: H-C Aasheim Received 8 November 1996; revised 17 December 1996; accepted 17 The PH domain is found in a large variety of proteins December 1996 involved in cellular signalling and cytoskeletal func- Identification of the homologue to the Lsc oncogene H-C Aasheim et al 1748 tions. It is believed that the function of the PH domain is to target the host protein to the cell membrane, by binding to lipids or proteins (Lemmon et al., 1996), to facilitate and regulate enzymatic activities. cDNA sequence and predicted protein structure of the sub1.5 cDNA sequence We have identi®ed and sequenced a novel cDNA encoding a putative protein with homology to two recently described members of the Dbl-family of oncogenes, Lbc and Lfc (Toksoz and Williams, 1994; Whitehead et al., 1995). A novel 3.4 kb cDNA was isolated from a human TPA-stimulated T cell cDNA library using a subtractive strategy recently described (Aasheim et al., 1994, 1996). The sequence revealed an open reading frame (ORF) of 875 amino acids, starting with an ATG codon at nucleotide 436 and terminating with a stop codon at nucleotide 3044 (data not shown, accession number Y09160). The ®rst ATG codon is in moderately good context for translation initiation with the sequence, CACCTCATGG as compared with the Kozak consensus sequence (XCXGCCATGG;Kozak, 1994). Recently, a mouse cDNA (Lsc, Whitehead et al., 1996) was published with 87% overall homology at the amino acid level to the sub1.5 sequence. The overall amino acid homol- ogy, the similar expression pattern and the transcript sizes strongly suggest that sub1.5 is the human homolog of the corresponding mouse gene. The mouse protein is denoted Lsc (Lbc's second cousin) and is suggested to be an oncogene (Whitehead et al., 1996). The Lsc cDNA was isolated based on the ability to induce strong oncogenic transformation when expressed in NIH3T3 ®broblast cells. The Lsc sequence is proposed to start at an ATG 53 amino acids upstream of the start of the sub1.5 sequence (Figure 1). Sub1.5 and Lsc diverge from nucleotide Figure 1 Comparison of the amino acid sequence of sub1.5 and number 334 in the sub1.5 sequence and 5' upstream. Lsc. The sequences were optimally aligned on the basis of identical residues (vertical lines). The DH-domain and the PH- Here, the two sequences do not show any homology to domain is localized from amino acid 359 to 710 in the sub1.5 each other. The sub1.5 sequence does not harbour an sequence ATG for translation start of the corresponding site where Lsc is proposed to start (data not shown). The sub1.5 cDNA encodes a protein with a predicted molecular weight of 97 kDa with several interesting transforming activity and the GEF activity of features. Thus, based on homology to previous known oncogenic Dbl. So far GEF activity has been shown genes, the predicted sub1.5 protein and Lsc show for Dbl (Hart et al., 1994), Ost (Horii et al., 1994), homology to the Dbl family of GEF's, and most Cdc24 (Zheng et al., 1994), Tiam-1 (Van Leeuwen et closely related to the Dbl family member Lbc, an al., 1995) and Lbc (Zheng et al., 1995), all exhibiting exchange factor with speci®city for Rho family exchange activity in vitro on Rho family GTPases. All GTPases (Toksoz and Williams, 1994; Zheng et al., DH family members, including sub1.5 and Lsc, also 1995). Like all Dbl family members, sub1.5 contains a possess a PH domain. PH domains are thought to PH domain in tandem with the DH-domain. Both anchore proteins to cell membranes, either through Sub1.5 and Lsc are distinguished from Lbc by an interaction with lipids or with proteins (Gibson et al., extended N-terminus (more than 400 amino acids) that 1994; Lemmon et al., 1996). Both sub1.5 and Lsc does not appear to be necessary for, or inhibitory to possess a PH domain. The transforming activity of cellular transformation (Whitehead et al., 1996). This both the Lfc and Lbc oncogene has been shown to be area does not align signi®cantly to any other proteins dependent of the PH domain (Whitehead et al., 1995, when performing GenBank searches. 1996) indicating that a PH-dependent recruitment of The DH-domain is found in a number of proteins Dbl family members to the plasma membrane may be a suspected to be involved in cell growth regulation.
Load more

Recommended publications
  • Differential Requirement for SUB1 in Chromosomal and Plasmid Double-Strand DNA Break Repair
    University of Massachusetts Medical School eScholarship@UMMS University of Massachusetts Medical School Faculty Publications 2013-03-12 Differential requirement for SUB1 in chromosomal and plasmid double-strand DNA break repair Lijian Yu University of Massachusetts Medical School Et al. Let us know how access to this document benefits ou.y Follow this and additional works at: https://escholarship.umassmed.edu/faculty_pubs Part of the Amino Acids, Peptides, and Proteins Commons, Biochemistry Commons, Enzymes and Coenzymes Commons, Genetic Phenomena Commons, Molecular Biology Commons, and the Molecular Genetics Commons Repository Citation Yu L, Volkert MR. (2013). Differential requirement for SUB1 in chromosomal and plasmid double-strand DNA break repair. University of Massachusetts Medical School Faculty Publications. https://doi.org/ 10.1371/journal.pone.0058015. Retrieved from https://escholarship.umassmed.edu/faculty_pubs/248 This material is brought to you by eScholarship@UMMS. It has been accepted for inclusion in University of Massachusetts Medical School Faculty Publications by an authorized administrator of eScholarship@UMMS. For more information, please contact [email protected]. Differential Requirement for SUB1 in Chromosomal and Plasmid Double-Strand DNA Break Repair Lijian Yu, Michael R. Volkert* Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America Abstract Non homologous end joining (NHEJ) is an important process that repairs double strand DNA breaks (DSBs) in eukaryotic cells. Cells defective in NHEJ are unable to join chromosomal breaks. Two different NHEJ assays are typically used to determine the efficiency of NHEJ. One requires NHEJ of linearized plasmid DNA transformed into the test organism; the other requires NHEJ of a single chromosomal break induced either by HO endonuclease or the I-SceI restriction enzyme.
    [Show full text]
  • Proteomic Analysis of the Mediator Complex Interactome in Saccharomyces Cerevisiae Received: 26 October 2016 Henriette Uthe, Jens T
    www.nature.com/scientificreports OPEN Proteomic Analysis of the Mediator Complex Interactome in Saccharomyces cerevisiae Received: 26 October 2016 Henriette Uthe, Jens T. Vanselow & Andreas Schlosser Accepted: 25 January 2017 Here we present the most comprehensive analysis of the yeast Mediator complex interactome to date. Published: 27 February 2017 Particularly gentle cell lysis and co-immunopurification conditions allowed us to preserve even transient protein-protein interactions and to comprehensively probe the molecular environment of the Mediator complex in the cell. Metabolic 15N-labeling thereby enabled stringent discrimination between bona fide interaction partners and nonspecifically captured proteins. Our data indicates a functional role for Mediator beyond transcription initiation. We identified a large number of Mediator-interacting proteins and protein complexes, such as RNA polymerase II, general transcription factors, a large number of transcriptional activators, the SAGA complex, chromatin remodeling complexes, histone chaperones, highly acetylated histones, as well as proteins playing a role in co-transcriptional processes, such as splicing, mRNA decapping and mRNA decay. Moreover, our data provides clear evidence, that the Mediator complex interacts not only with RNA polymerase II, but also with RNA polymerases I and III, and indicates a functional role of the Mediator complex in rRNA processing and ribosome biogenesis. The Mediator complex is an essential coactivator of eukaryotic transcription. Its major function is to communi- cate regulatory signals from gene-specific transcription factors upstream of the transcription start site to RNA Polymerase II (Pol II) and to promote activator-dependent assembly and stabilization of the preinitiation complex (PIC)1–3. The yeast Mediator complex is composed of 25 subunits and forms four distinct modules: the head, the middle, and the tail module, in addition to the four-subunit CDK8 kinase module (CKM), which can reversibly associate with the 21-subunit Mediator complex.
    [Show full text]
  • Aneuploidy: Using Genetic Instability to Preserve a Haploid Genome?
    Health Science Campus FINAL APPROVAL OF DISSERTATION Doctor of Philosophy in Biomedical Science (Cancer Biology) Aneuploidy: Using genetic instability to preserve a haploid genome? Submitted by: Ramona Ramdath In partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biomedical Science Examination Committee Signature/Date Major Advisor: David Allison, M.D., Ph.D. Academic James Trempe, Ph.D. Advisory Committee: David Giovanucci, Ph.D. Randall Ruch, Ph.D. Ronald Mellgren, Ph.D. Senior Associate Dean College of Graduate Studies Michael S. Bisesi, Ph.D. Date of Defense: April 10, 2009 Aneuploidy: Using genetic instability to preserve a haploid genome? Ramona Ramdath University of Toledo, Health Science Campus 2009 Dedication I dedicate this dissertation to my grandfather who died of lung cancer two years ago, but who always instilled in us the value and importance of education. And to my mom and sister, both of whom have been pillars of support and stimulating conversations. To my sister, Rehanna, especially- I hope this inspires you to achieve all that you want to in life, academically and otherwise. ii Acknowledgements As we go through these academic journeys, there are so many along the way that make an impact not only on our work, but on our lives as well, and I would like to say a heartfelt thank you to all of those people: My Committee members- Dr. James Trempe, Dr. David Giovanucchi, Dr. Ronald Mellgren and Dr. Randall Ruch for their guidance, suggestions, support and confidence in me. My major advisor- Dr. David Allison, for his constructive criticism and positive reinforcement.
    [Show full text]
  • Disruption of Transcriptional Coactivator Sub1 Leads to Genome-Wide Re-Distribution of Clustered Mutations Induced by APOBEC in Active Yeast Genes
    RESEARCH ARTICLE Disruption of Transcriptional Coactivator Sub1 Leads to Genome-Wide Re-distribution of Clustered Mutations Induced by APOBEC in Active Yeast Genes Artem G. Lada1☯*, Sergei F. Kliver2☯, Alok Dhar3, Dmitrii E. Polev4, Alexey E. Masharsky4, Igor B. Rogozin5,6,7, Youri I. Pavlov1* 1 Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska, United States of America, 2 Department of Genetics and Biotechnology, Saint Petersburg State University, St. Petersburg, Russia, 3 Department of Genetics, Cell Biology and Anatomy, and Munroe-Meyer Institute, University of Nebraska Medical Center, Omaha, Nebraska, United States of America, 4 Research Resource Center for Molecular and Cell Technologies, Research Park, St. Petersburg State University, St. Petersburg, Russia, 5 National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, Maryland, United States of America, 6 Institute of Cytology and Genetics, Novosibirsk, Russia, 7 Novosibirsk State University, Novosibirsk, Russia OPEN ACCESS ☯ These authors contributed equally to this work. * Citation: Lada AG, Kliver SF, Dhar A, Polev DE, [email protected] (AGL); [email protected] (YIP) Masharsky AE, Rogozin IB, et al. (2015) Disruption of Transcriptional Coactivator Sub1 Leads to Genome- Wide Re-distribution of Clustered Mutations Induced Abstract by APOBEC in Active Yeast Genes. PLoS Genet 11 (5): e1005217. doi:10.1371/journal.pgen.1005217 Mutations in genomes of species are frequently distributed non-randomly, resulting in muta- Received: February 3, 2015 tion clusters, including recently discovered kataegis in tumors. DNA editing deaminases Accepted: April 13, 2015 play the prominent role in the etiology of these mutations.
    [Show full text]
  • Small-Molecule G-Quadruplex Stabilizers Reveal a Novel Pathway
    RESEARCH ARTICLE Small-molecule G-quadruplex stabilizers reveal a novel pathway of autophagy regulation in neurons Jose F Moruno-Manchon1, Pauline Lejault2, Yaoxuan Wang1, Brenna McCauley3, Pedram Honarpisheh4,5, Diego A Morales Scheihing4, Shivani Singh6, Weiwei Dang3, Nayun Kim6, Akihiko Urayama4,5, Liang Zhu7,8, David Monchaud2, Louise D McCullough4,5, Andrey S Tsvetkov1,5,9* 1Department of Neurobiology and Anatomy, The University of Texas McGovern Medical School at Houston, Houston, United States; 2Institut de Chimie Mole´culaire (ICMUB), UBFC Dijon, CNRS UMR6302, Dijon, France; 3Huffington Center on Aging, Baylor College of Medicine, Houston, United States; 4Department of Neurology, The University of Texas McGovern Medical School at Houston, Houston, United States; 5The University of Texas Graduate School of Biomedical Sciences, Houston, United States; 6Department of Microbiology and Molecular Genetics, The University of Texas McGovern Medical School at Houston, Houston, United States; 7Biostatistics and Epidemiology Research Design Core Center for Clinical and Translational Sciences, The University of Texas McGovern Medical School at Houston, Houston, United States; 8Department of Internal Medicine, The University of Texas McGovern Medical School at Houston, Houston, United States; 9UTHealth Consortium on Aging, The University of Texas McGovern Medical School at Houston, Houston, United States Abstract Guanine-rich DNA sequences can fold into four-stranded G-quadruplex (G4-DNA) *For correspondence: structures. G4-DNA regulates replication and transcription, at least in cancer cells. Here, we [email protected] demonstrate that, in neurons, pharmacologically stabilizing G4-DNA with G4 ligands strongly downregulates the Atg7 gene. Atg7 is a critical gene for the initiation of autophagy that exhibits Competing interests: The decreased transcription with aging.
    [Show full text]
  • Product Size GOT1 P00504 F CAAGCTGT
    Table S1. List of primer sequences for RT-qPCR. Gene Product Uniprot ID F/R Sequence(5’-3’) name size GOT1 P00504 F CAAGCTGTCAAGCTGCTGTC 71 R CGTGGAGGAAAGCTAGCAAC OGDHL E1BTL0 F CCCTTCTCACTTGGAAGCAG 81 R CCTGCAGTATCCCCTCGATA UGT2A1 F1NMB3 F GGAGCAAAGCACTTGAGACC 93 R GGCTGCACAGATGAACAAGA GART P21872 F GGAGATGGCTCGGACATTTA 90 R TTCTGCACATCCTTGAGCAC GSTT1L E1BUB6 F GTGCTACCGAGGAGCTGAAC 105 R CTACGAGGTCTGCCAAGGAG IARS Q5ZKA2 F GACAGGTTTCCTGGCATTGT 148 R GGGCTTGATGAACAACACCT RARS Q5ZM11 F TCATTGCTCACCTGCAAGAC 146 R CAGCACCACACATTGGTAGG GSS F1NLE4 F ACTGGATGTGGGTGAAGAGG 89 R CTCCTTCTCGCTGTGGTTTC CYP2D6 F1NJG4 F AGGAGAAAGGAGGCAGAAGC 113 R TGTTGCTCCAAGATGACAGC GAPDH P00356 F GACGTGCAGCAGGAACACTA 112 R CTTGGACTTTGCCAGAGAGG Table S2. List of differentially expressed proteins during chronic heat stress. score name Description MW PI CC CH Down regulated by chronic heat stress A2M Uncharacterized protein 158 1 0.35 6.62 A2ML4 Uncharacterized protein 163 1 0.09 6.37 ABCA8 Uncharacterized protein 185 1 0.43 7.08 ABCB1 Uncharacterized protein 152 1 0.47 8.43 ACOX2 Cluster of Acyl-coenzyme A oxidase 75 1 0.21 8 ACTN1 Alpha-actinin-1 102 1 0.37 5.55 ALDOC Cluster of Fructose-bisphosphate aldolase 39 1 0.5 6.64 AMDHD1 Cluster of Uncharacterized protein 37 1 0.04 6.76 AMT Aminomethyltransferase, mitochondrial 42 1 0.29 9.14 AP1B1 AP complex subunit beta 103 1 0.15 5.16 APOA1BP NAD(P)H-hydrate epimerase 32 1 0.4 8.62 ARPC1A Actin-related protein 2/3 complex subunit 42 1 0.34 8.31 ASS1 Argininosuccinate synthase 47 1 0.04 6.67 ATP2A2 Cluster of Calcium-transporting
    [Show full text]
  • 393LN V 393P 344SQ V 393P Probe Set Entrez Gene
    393LN v 393P 344SQ v 393P Entrez fold fold probe set Gene Gene Symbol Gene cluster Gene Title p-value change p-value change chemokine (C-C motif) ligand 21b /// chemokine (C-C motif) ligand 21a /// chemokine (C-C motif) ligand 21c 1419426_s_at 18829 /// Ccl21b /// Ccl2 1 - up 393 LN only (leucine) 0.0047 9.199837 0.45212 6.847887 nuclear factor of activated T-cells, cytoplasmic, calcineurin- 1447085_s_at 18018 Nfatc1 1 - up 393 LN only dependent 1 0.009048 12.065 0.13718 4.81 RIKEN cDNA 1453647_at 78668 9530059J11Rik1 - up 393 LN only 9530059J11 gene 0.002208 5.482897 0.27642 3.45171 transient receptor potential cation channel, subfamily 1457164_at 277328 Trpa1 1 - up 393 LN only A, member 1 0.000111 9.180344 0.01771 3.048114 regulating synaptic membrane 1422809_at 116838 Rims2 1 - up 393 LN only exocytosis 2 0.001891 8.560424 0.13159 2.980501 glial cell line derived neurotrophic factor family receptor alpha 1433716_x_at 14586 Gfra2 1 - up 393 LN only 2 0.006868 30.88736 0.01066 2.811211 1446936_at --- --- 1 - up 393 LN only --- 0.007695 6.373955 0.11733 2.480287 zinc finger protein 1438742_at 320683 Zfp629 1 - up 393 LN only 629 0.002644 5.231855 0.38124 2.377016 phospholipase A2, 1426019_at 18786 Plaa 1 - up 393 LN only activating protein 0.008657 6.2364 0.12336 2.262117 1445314_at 14009 Etv1 1 - up 393 LN only ets variant gene 1 0.007224 3.643646 0.36434 2.01989 ciliary rootlet coiled- 1427338_at 230872 Crocc 1 - up 393 LN only coil, rootletin 0.002482 7.783242 0.49977 1.794171 expressed sequence 1436585_at 99463 BB182297 1 - up 393
    [Show full text]
  • Datasheet: VPA00736KT Product Details
    Datasheet: VPA00736KT Description: PC4 ANTIBODY WITH CONTROL LYSATE Specificity: PC4 Format: Purified Product Type: PrecisionAb™ Polyclonal Isotype: Polyclonal IgG Quantity: 2 Westerns Product Details Applications This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit www.bio-rad-antibodies.com/protocols. Yes No Not Determined Suggested Dilution Western Blotting 1/1000 PrecisionAb antibodies have been extensively validated for the western blot application. The antibody has been validated at the suggested dilution. Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Further optimization may be required dependant on sample type. Target Species Human Product Form Purified IgG - liquid Preparation 20μl Rabbit polyclonal antibody purified by affinity chromatography on Protein A Buffer Solution Phosphate buffered saline Preservative 0.09% Sodium Azide Stabilisers 2% Sucrose Immunogen Synthetic peptide encompassing part of the middle region of human PC4 External Database Links UniProt: P53999 Related reagents Entrez Gene: 10923 SUB1 Related reagents Synonyms PC4, RPO2TC1 Specificity Rabbit anti Human PC4 antibody recognizes the activated RNA polymerase II transcriptional coactivator p15, also known as activated RNA polymerase II transcription cofactor 4, or positive cofactor 4. Page 1 of 2 This protein forms a homodimer and is aneneral coactivator that functions cooperatively with TAFs and mediates functional interactions between upstream activators and the general transcriptional machinery. It also may be involved in stabilizing the multiprotein transcription complex.
    [Show full text]
  • Genome-Wide Location Analysis Reveals a Role for Sub1 in RNA Polymerase III Transcription
    Genome-wide location analysis reveals a role for Sub1 in RNA polymerase III transcription Arounie Taveneta, Audrey Suleaua,1,Ge´ raldine Dubreuila, Roberto Ferrarib,2,Ce´ cile Ducrota, Magali Michauta, Jean-Christophe Audea, Giorgio Diecib, Olivier Lefebvrea, Christine Conesaa,3, and Joe¨l Ackera,3 aCommissariat a`l’Energie Atomique, Institut de Biologie et de Technologies de Saclay, F-91101 Gif sur Yvette, France; and bDipartimento di Biochimica e Biologia Molecolare, Universita`degli Studi di Parma, 43100 Parma, Italy Edited by Robert G. Roeder, The Rockefeller University, New York, NY, and approved June 30, 2009 (received for review January 8, 2009) Human PC4 and the yeast ortholog Sub1 have multiple functions in that we used in a previous work to analyze the genomic location of RNA polymerase II transcription. Genome-wide mapping revealed the Pol III transcription machinery (15). A defined number of loci that Sub1 is present on Pol III-transcribed genes. Sub1 was found were significantly enriched (991 loci with a P value Ͻ0.01) under to interact with components of the Pol III transcription system and active growth conditions. Approximately one-fourth of the en- to stimulate the initiation and reinitiation steps in a system riched loci were located within ORF; the others corresponded to reconstituted with all recombinant factors. Sub1 was required for intergenic regions and to genes encoding nontranslated RNAs. We optimal Pol III gene transcription in exponentially growing cells. noted that the ACT1, PMA1, PYK1, ADH1, and snoRNA genes previously identified as DNA targets of Sub1 by ChIP and PCR Chip chip ͉ PC4 ͉ reinitiation ͉ TFIIIB amplification (10, 16, 17) were indeed enriched in our data.
    [Show full text]
  • Autocrine IFN Signaling Inducing Profibrotic Fibroblast Responses By
    Downloaded from http://www.jimmunol.org/ by guest on September 23, 2021 Inducing is online at: average * The Journal of Immunology , 11 of which you can access for free at: 2013; 191:2956-2966; Prepublished online 16 from submission to initial decision 4 weeks from acceptance to publication August 2013; doi: 10.4049/jimmunol.1300376 http://www.jimmunol.org/content/191/6/2956 A Synthetic TLR3 Ligand Mitigates Profibrotic Fibroblast Responses by Autocrine IFN Signaling Feng Fang, Kohtaro Ooka, Xiaoyong Sun, Ruchi Shah, Swati Bhattacharyya, Jun Wei and John Varga J Immunol cites 49 articles Submit online. Every submission reviewed by practicing scientists ? is published twice each month by Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts http://jimmunol.org/subscription Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html http://www.jimmunol.org/content/suppl/2013/08/20/jimmunol.130037 6.DC1 This article http://www.jimmunol.org/content/191/6/2956.full#ref-list-1 Information about subscribing to The JI No Triage! Fast Publication! Rapid Reviews! 30 days* Why • • • Material References Permissions Email Alerts Subscription Supplementary The Journal of Immunology The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2013 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. This information is current as of September 23, 2021. The Journal of Immunology A Synthetic TLR3 Ligand Mitigates Profibrotic Fibroblast Responses by Inducing Autocrine IFN Signaling Feng Fang,* Kohtaro Ooka,* Xiaoyong Sun,† Ruchi Shah,* Swati Bhattacharyya,* Jun Wei,* and John Varga* Activation of TLR3 by exogenous microbial ligands or endogenous injury-associated ligands leads to production of type I IFN.
    [Show full text]
  • Altered Transcriptome-Proteome Coupling Indicates Aberrant Proteostasis in Parkinson’S Disease Fiona Dick1,2, Ole-Bjørn Tysnes1,2, Guido Werner Alves3,4, Gonzalo S
    medRxiv preprint doi: https://doi.org/10.1101/2021.03.18.21253875; this version posted March 20, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license . Altered transcriptome-proteome coupling indicates aberrant proteostasis in Parkinson's disease Fiona Dick1,2, Ole-Bjørn Tysnes1,2, Guido Werner Alves3,4, Gonzalo S. Nido1,2, Charalampos Tzoulis1,2* 1 Neuro-SysMed, Department of Neurology, Haukeland University Hospital, Bergen, Norway 2 Department of Clinical Medicine, University of Bergen, Bergen, Norway 3 The Norwegian Center for Movement Disorders and Department of Neurology, Stavanger University Hospital, Stavanger, Norway 4 Department of Mathematics and Natural Sciences, University of Stavanger, Stavanger, Norway * [email protected] Abstract The correlation between mRNA and protein levels has been shown to decline in the ageing brain, possibly reflecting age-dependent changes in the proteostasis. It is thought that impaired proteostasis may be implicated in the pathogenesis of Parkinson's disease (PD), but evidence derived from the patient brain is currently limited. Here, we hypothesized that if impaired proteostasis occurs in PD, this should be reflected in the form of altered correlation between transcriptome and proteome compared to healthy ageing. To test this hypothesis, we integrated transcriptomic data with proteomics from prefrontal cortex tissue of 17 PD patients and 11 demographically matched healthy controls and assessed gene-specific correlations between RNA and protein level. To control for the effects of ageing, brain samples from 4 infants were included in the analyses.
    [Show full text]
  • Substitution of Tryptophan 89 with Tyrosine Switches the DNA Binding
    OPEN Substitution of tryptophan 89 with SUBJECT AREAS: tyrosine switches the DNA binding mode MICROBIOLOGY X-RAY CRYSTALLOGRAPHY of PC4 Jinguang Huang1,2,3*, Yanxiang Zhao2*, Huaian Liu2, Dan Huang2, Xiankun Cheng2, Wensheng Zhao1,2, Received Ian A. Taylor4, Junfeng Liu2 & You-Liang Peng1,2 27 October 2014 Accepted 1State key Laboratory of Agrobiotechnology, China Agricultural University, No2 Yunamingyuanxilu, Beijing, 100193, China, 4 February 2015 2MOA Key Laboratory of Plant Pathology, China Agricultural University, No2 Yunamingyuanxilu, Beijing, 100193, China, 3College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao, Shandong, 266109, China, 4Molecular Published Structure, MRC-NIMR, The Ridgeway, London, NW7 1AA, United Kingdom. 5 March 2015 PC4, a well-known general transcription cofactor, has multiple functions in transcription and DNA repair. Correspondence and Residue W89, is engaged in stacking interactions with DNA in PC4, but substituted by tyrosine in some PC4 orthologous proteins. In order to understand the consequences and reveal the molecular details of this requests for materials substitution we have determined the crystal structures of the PC4 orthologue MoSub1 and a PC4 W89Y should be addressed to mutant in complex with DNA. In the structure of MoSub1-DNA complex, Y74 interacts directly with a J.L. ([email protected]) single nucleotide of oligo DNA. By comparison, the equivalent residue, W89 in wild type PC4 interacts with two nucleotides and the base of the second nucleotide has distinct orientation relative to that of the first one. A hydrophobic patch around W89 that favours interaction with two nucleotides is not formed in the PC4 * These authors W89Y mutant.
    [Show full text]