DOCSLIB.ORG

The Flightless I Protein

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Flightless I Protein RESEARCH ARTICLE 549 The flightless I protein colocalizes with actin- and microtubule-based structures in motile Swiss 3T3 fibroblasts: evidence for the involvement of PI 3-kinase and Ras-related small GTPases Deborah A. Davy1,*, Hugh D. Campbell2, Shelley Fountain2, Danielle de Jong2 and Michael F. Crouch1,* 1Molecular Signalling Group, Division of Neuroscience, John Curtin School of Medical Research and 2Molecular Genetics and Evolution Group and Centre for Molecular Structure and Function, Research School of Biological Sciences, Australian National University, Canberra, Australia 2600 *Authors for correspondence (email: [email protected]; [email protected]) Accepted 17 November 2000 Journal of Cell Science 114, 549-562 © The Company of Biologists Ltd SUMMARY The flightless I protein contains an actin-binding domain protein colocalizes with β-tubulin- and actin-based with homology to the gelsolin family and is likely to be structures. Members of the small GTPase family, also involved in actin cytoskeletal rearrangements. It has been implicated in cytoskeletal control, were found to colocalize suggested that this protein is involved in linking the with flightless I in migrating Swiss 3T3 fibroblasts. cytoskeletal network with signal transduction pathways. LY294002, a specific inhibitor of PI 3-kinase, inhibits the We have developed antibodies directed toward the leucine translocation of flightless I to actin-based structures. Our rich repeat and gelsolin-like domains of the human and results suggest that PI 3-kinase and the small GTPases, mouse homologues of flightless I that specifically recognize Ras, RhoA and Cdc42 may be part of a common functional expressed and endogenous forms of the protein. We have pathway involved in Fliih-mediated cytoskeletal regulation. also constructed a flightless I-enhanced green fluorescent Functionally, we suggest that flightless I may act to prepare fusion vector and used this to examine the localization of actin filaments or provide factors required for cytoskeletal the expressed protein in Swiss 3T3 fibroblasts. The rearrangements necessary for cell migration and/or flightless I protein localizes predominantly to the nucleus adhesion. and translocates to the cytoplasm following serum stimulation. In cells stimulated to migrate, the flightless I Key words: Flightless I, Fliih, Cytoskeleton, GTPase, Actin, Tubulin INTRODUCTION moderate and severe fliI mutants indicate a possible role for the fliI protein in regulating actin cytoskeletal rearrangements. The Drosophila melanogaster flightless I (fliI) gene encodes a A 380 amino acid leucine-rich repeat region (LRR) (Kobe 1,256 amino acid protein with a predicted molecular mass of and Deisenhofer, 1995a), characterizes the N-terminal domain 143,672 Da (Campbell et al., 1993). It has highly conserved of the fliI protein. This motif is thought to play a role in specific homologues in Caenorhabditis elegans (49% amino acid protein-protein interactions, that in some cases are responsible sequence identity to Drosophila), mouse (95% identity to for a role in signal transduction (Kobe and Deisenhofer, human) (Campbell et al., 2000) and human (58% identity to 1995b). This amphipathic beta sheet/alpha helix motif (Kobe Drosophila) (Campbell et al., 1993; Campbell et al., 1997). and Deisenhofer, 1994; Kobe and Deisenhofer, 1995b) was first Northern analysis of human FLII mRNA expression patterns observed in the α2-glycoprotein (Takahashi et al., 1985) and showed the highest level in skeletal muscle, but the gene was has since been identified in hormone receptors (McFarland et expressed at varying levels in all tissues examined (Campbell al., 1989), certain enzymes (Tan et al., 1990), tyrosine kinase et al., 1997). Viable mutations of the D. melanogaster fliI locus receptors (Martin-Zanca et al., 1989) and molecules cause ultrastructural defects in the indirect flight muscles responsible for cell adhesion (Hashimoto et al., 1988). A (Deak et al., 1982; Miklos and de Couet, 1990), with frayed variety of LRR functions have been characterized (Kobe and and disorganized myofibrils and Z bands that appear abnormal Deisenhofer, 1994), including the LRR region in yeast or absent. Null alleles of fliI are homozygous lethal (Perrimon adenylate cyclase which is known to interact with Ras (Suzuki et al., 1989). In contrast to normal embryonic development et al., 1990). (Miller and Kiehart, 1995), mutant embryos have an irregular C-terminal to the LRR region lie two large duplicated cytoskeleton and arrangement of nuclei at the cell cortex, domains each characterized by three tandemly repeated followed by poorly coordinated membrane invaginations sequences of between 125 and 150 amino acids (Campbell et (Kajava et al., 1995; Straub et al., 1996). These developmental al., 1993; Way and Weeds, 1988). This region shows significant events require an intact cytoskeleton. The phenotypes of homology to members of the gelsolin-family of actin-binding 550 JOURNAL OF CELL SCIENCE 114 (3) proteins (ABPs) (Campbell et al., 1997; Kwiatkowski et al., 50 µg of peptide was incubated with 50 µl of the antibody for one 1986; Hartwig and Kwiatkowski, 1991). The gelsolin family hour at room temperature. This preparation was then used in the same members are ABPs that can bind monomeric actin subunits manner as antibodies that were not blocked. promoting polymerization (Janmey et al., 1985; Yin, 1986; Construction of a plasmid containing the complete human Kwiatkowski et al., 1989). Members of this family can cap FLII coding region (Lind et al., 1987) and/or sever actin filaments (Kwiatkowski The previously reported human FLII cDNA (phfli2) (Campbell et al., et al., 1989; Arora and McCulloch, 1996; Chaponnier et al., 1993) was almost full length, but was missing a part of the ATG 1986). ABPs are important for actin cytoskeletal rearrangements initiation codon. A human EST clone, Image Consortium clone and are regulated by micromolar concentrations of calcium 135082 (GenBank accession no. R33910), was previously identified (Janmey et al., 1985; Pope et al., 1995; Yin et al., 1988; Young as containing the ATG initiation codon and 37 bp of 5′ untranslated et al., 1994) and phosphoinositide binding (Janmey and Stossel, sequence (Campbell et al., 1997). However, this EST clone contained 1989; Lassing and Lindberg, 1985; Lu et al., 1996). an insert of only 800 bp. Clone 135082 plasmid DNA was digested The cytoskeletal network plays important roles in the with XhoI and end repaired with T4 DNA polymerase. The DNA was maintenance of cell shape, the transport and anchoring of digested with BspEI and the 300 bp XhoI-BspEI fragment was gel cellular components involved in cellular adhesion, and purified. Plasmid phfli2 was digested with EcoRV and BspEI, migration. While significant progress has been made in dephosphorylated with calf intestinal alkaline phosphatase, and the 6.8 kb fragment was gel purified. These two fragments were ligated identifying the biochemical signals that are involved in together to yield phfli2FL. The insert in phfli2FL was sequenced from regulating the cytoskeleton, the relationship between them and both ends by dye primer sequencing (ABI Prism) using the –21 M13 the impact on cellular functions are not well defined. Recent forward and reverse primers, confirming the expected structure. The evidence has shown that members of the Ras-related GTPase 5′ sequence of the phfli2FL cDNA up to and across the BspEI site was family are responsible for the regulation of a variety of actin- identical to the previously determined cDNA and genomic sequences based structures (Allen et al., 1997; Nobes and Hall, 1995; (Campbell et al., 1993; Campbell et al., 1997). Tapon and Hall, 1997; Rodriguez-Viciana et al., 1997). In addition to this, signaling pathways involved in the regulation In vitro transcription/translation of phfli2FL of proteins that influence cytoskeletal dynamics are The transcription/translation of phfli2FL plasmid DNA was achieved increasingly seen to involve phosphoinositide 3-kinase (PI 3- using the TNT T7 Quick Coupled Translation/Transcription System (Promega). [35S]methionine was used to produce a radioactive form kinase) (Derman et al., 1997; Rodriguez-Viciana et al., 1997; of protein that can, when appropriately prepared, be visualized using Johanson et al., 1999; Hill et al., 2000). autoradiography. A 5µl aliquot from each reaction was added to SDS- The work presented here describes the development of PAGE sample buffer (300 mM Tris, 50 mM DTT, 15% glycerol, 2% flightless I-specific antibodies and the identification and SDS, bromophenol blue) plus 15 mg/ml dithiothreitol (DTT), or 1 ml examination of the subcellular distribution of the mouse of lysis buffer containing inhibitors (see Cell Harvest methods). flightless I protein (Fliih) protein in quiescent and serum- Samples containing lysis buffer and 5 µl of transcription/translation stimulated Swiss 3T3 fibroblasts. We also examine potential reaction were heated for 30 minutes at 70°C and precleared by interacting proteins and regulatory molecules for Fliih. The incubation with Protein A-Sepharose beads only for one hour at 4°C. FliL or FliG antibodies (1:50) were added and samples were incubated results of our experiments imply a role for flightless I in ° the regulation of cytoskeletal rearrangements involved in overnight at 4 C on a rotating mixer. Protein A-Sepharose beads were hydrated with Tris buffered saline (TBS: 50 mM Tris-HCl, pH 7.4, cytokinesis and cell migration. 0.2 M NaCl) and 5 mg (w/v) was added to each sample. The samples were mixed for 1 hour at 4°C. The beads were pelleted by centrifugation for 45 seconds at 17,400 g, and the supernatant MATERIALS AND METHODS discarded. The Protein A-Sepharose-antibody-antigen pellets were rinsed 3 times with 1 ml of TBS. SDS-PAGE sample buffer was added Immunization with flightless I-specific-peptide L to each pellet and samples were analyzed by SDS-PAGE. The SDS- The cDNA sequences of flightless I and homologues have been PAGE gel was dried prior to autoradiography. determined (Campbell et al., 1993) (D. melanogaster GenBank accession no. U01182, C. elegans GenBank U01183 and Homo Bacterial expression of flightless I sapiens GenBank U01184; U80184).
Load more

Recommended publications
  • October 2020
    PCORI Health Care Horizon Scanning System: Horizon Scanning COVID-19 Supplement Status Report Volume 1, Issue 2 Prepared for: Patient-Centered Outcomes Research Institute 1828 L St., NW, Suite 900 Washington, DC 20036 Contract No. MSA-HORIZSCAN-ECRI-ENG-2018.7.12 Prepared by: ECRI 5200 Butler Pike Plymouth Meeting, PA 19462 Investigators: Randy Hulshizer, MA, MS Marcus Lynch, PhD, MBA Jennifer De Lurio, MS Brian Wilkinson, MA Damian Carlson, MS Christian Cuevas, PhD Andrea Druga, MSPAS, PA-C Misha Mehta, MS Prital Patel, MPH Donna Beales, MLIS Eloise DeHaan, BS Eileen Erinoff, MSLIS Cassia Hulshizer, AS Madison Kimball, MS Maria Middleton, MPH Diane Robertson, BA Kelley Tipton, MPH Rosemary Walker, MLIS Karen Schoelles, MD, SM October 2020 Statement of Funding and Purpose This report incorporates data collected during implementation of the Patient-Centered Outcomes Research Institute (PCORI) Health Care Horizon Scanning System COVID-19 Supplement, operated by ECRI under contract to PCORI, Washington, DC (Contract No. MSA- HORIZSCAN-ECRI-ENG-2018.7.12). The findings and conclusions in this document are those of the authors, who are responsible for its content. No statement in this report should be construed as an official position of PCORI. An innovation that potentially meets inclusion criteria might not appear in this report simply because the horizon scanning system has not yet detected it or it does not yet meet inclusion criteria outlined in the PCORI Health Care Horizon Scanning System: Horizon Scanning Protocol and Operations Manual COVID-19 Supplement. Inclusion or absence of innovations in the horizon scanning reports will change over time as new information is collected; therefore, inclusion or absence should not be construed as either an endorsement or rejection of specific interventions.
    [Show full text]
  • Genes in a Refined Smith-Magenis Syndrome Critical Deletion Interval on Chromosome 17P11.2 and the Syntenic Region of the Mouse
    Downloaded from genome.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press Article Genes in a Refined Smith-Magenis Syndrome Critical Deletion Interval on Chromosome 17p11.2 and the Syntenic Region of the Mouse Weimin Bi,1,6 Jiong Yan,1,6 Paweł Stankiewicz,1 Sung-Sup Park,1,7 Katherina Walz,1 Cornelius F. Boerkoel,1 Lorraine Potocki,1,3 Lisa G. Shaffer,1 Koen Devriendt,4 Małgorzata J.M. Nowaczyk,5 Ken Inoue,1 and James R. Lupski1,2,3,8 Departments of 1Molecular & Human Genetics, 2Pediatrics, Baylor College of Medicine, 3Texas Children’s Hospital, Houston, Texas 77030, USA; 4Centre for Human Genetics, University Hospital Gasthuisberg, Catholic University of Leuven, B-3000 Leuven, Belgium; 5Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario L8S 4J9, Canada Smith-Magenis syndrome (SMS) is a multiple congenital anomaly/mental retardation syndrome associated with behavioral abnormalities and sleep disturbance. Most patients have the same ∼4 Mb interstitial genomic deletion within chromosome 17p11.2. To investigate the molecular bases of the SMS phenotype, we constructed BAC/PAC contigs covering the SMS common deletion interval and its syntenic region on mouse chromosome 11. Comparative genome analysis reveals the absence of all three ∼200-kb SMS-REP low-copy repeats in the mouse and indicates that the evolution of SMS-REPs was accompanied by transposition of adjacent genes. Physical and genetic map comparisons in humans reveal reduced recombination in both sexes. Moreover, by examining the deleted regions in SMS patients with unusual-sized deletions, we refined the minimal Smith-Magenis critical region (SMCR) to an ∼1.1-Mb genomic interval that is syntenic to an ∼1.0-Mb region in the mouse.
    [Show full text]
  • Androgen Receptor Interacting Proteins and Coregulators Table
    ANDROGEN RECEPTOR INTERACTING PROTEINS AND COREGULATORS TABLE Compiled by: Lenore K. Beitel, Ph.D. Lady Davis Institute for Medical Research 3755 Cote Ste Catherine Rd, Montreal, Quebec H3T 1E2 Canada Telephone: 514-340-8260 Fax: 514-340-7502 E-Mail: [email protected] Internet: http://androgendb.mcgill.ca Date of this version: 2010-08-03 (includes articles published as of 2009-12-31) Table Legend: Gene: Official symbol with hyperlink to NCBI Entrez Gene entry Protein: Protein name Preferred Name: NCBI Entrez Gene preferred name and alternate names Function: General protein function, categorized as in Heemers HV and Tindall DJ. Endocrine Reviews 28: 778-808, 2007. Coregulator: CoA, coactivator; coR, corepressor; -, not reported/no effect Interactn: Type of interaction. Direct, interacts directly with androgen receptor (AR); indirect, indirect interaction; -, not reported Domain: Interacts with specified AR domain. FL-AR, full-length AR; NTD, N-terminal domain; DBD, DNA-binding domain; h, hinge; LBD, ligand-binding domain; C-term, C-terminal; -, not reported References: Selected references with hyperlink to PubMed abstract. Note: Due to space limitations, all references for each AR-interacting protein/coregulator could not be cited. The reader is advised to consult PubMed for additional references. Also known as: Alternate gene names Gene Protein Preferred Name Function Coregulator Interactn Domain References Also known as AATF AATF/Che-1 apoptosis cell cycle coA direct FL-AR Leister P et al. Signal Transduction 3:17-25, 2003 DED; CHE1; antagonizing regulator Burgdorf S et al. J Biol Chem 279:17524-17534, 2004 CHE-1; AATF transcription factor ACTB actin, beta actin, cytoplasmic 1; cytoskeletal coA - - Ting HJ et al.
    [Show full text]
  • 1 Supporting Information for a Microrna Network Regulates
    Supporting Information for A microRNA Network Regulates Expression and Biosynthesis of CFTR and CFTR-ΔF508 Shyam Ramachandrana,b, Philip H. Karpc, Peng Jiangc, Lynda S. Ostedgaardc, Amy E. Walza, John T. Fishere, Shaf Keshavjeeh, Kim A. Lennoxi, Ashley M. Jacobii, Scott D. Rosei, Mark A. Behlkei, Michael J. Welshb,c,d,g, Yi Xingb,c,f, Paul B. McCray Jr.a,b,c Author Affiliations: Department of Pediatricsa, Interdisciplinary Program in Geneticsb, Departments of Internal Medicinec, Molecular Physiology and Biophysicsd, Anatomy and Cell Biologye, Biomedical Engineeringf, Howard Hughes Medical Instituteg, Carver College of Medicine, University of Iowa, Iowa City, IA-52242 Division of Thoracic Surgeryh, Toronto General Hospital, University Health Network, University of Toronto, Toronto, Canada-M5G 2C4 Integrated DNA Technologiesi, Coralville, IA-52241 To whom correspondence should be addressed: Email: [email protected] (M.J.W.); yi- [email protected] (Y.X.); Email: [email protected] (P.B.M.) This PDF file includes: Materials and Methods References Fig. S1. miR-138 regulates SIN3A in a dose-dependent and site-specific manner. Fig. S2. miR-138 regulates endogenous SIN3A protein expression. Fig. S3. miR-138 regulates endogenous CFTR protein expression in Calu-3 cells. Fig. S4. miR-138 regulates endogenous CFTR protein expression in primary human airway epithelia. Fig. S5. miR-138 regulates CFTR expression in HeLa cells. Fig. S6. miR-138 regulates CFTR expression in HEK293T cells. Fig. S7. HeLa cells exhibit CFTR channel activity. Fig. S8. miR-138 improves CFTR processing. Fig. S9. miR-138 improves CFTR-ΔF508 processing. Fig. S10. SIN3A inhibition yields partial rescue of Cl- transport in CF epithelia.
    [Show full text]
  • Supplementary Materials
    Supplementary materials Supplementary Table S1: MGNC compound library Ingredien Molecule Caco- Mol ID MW AlogP OB (%) BBB DL FASA- HL t Name Name 2 shengdi MOL012254 campesterol 400.8 7.63 37.58 1.34 0.98 0.7 0.21 20.2 shengdi MOL000519 coniferin 314.4 3.16 31.11 0.42 -0.2 0.3 0.27 74.6 beta- shengdi MOL000359 414.8 8.08 36.91 1.32 0.99 0.8 0.23 20.2 sitosterol pachymic shengdi MOL000289 528.9 6.54 33.63 0.1 -0.6 0.8 0 9.27 acid Poricoic acid shengdi MOL000291 484.7 5.64 30.52 -0.08 -0.9 0.8 0 8.67 B Chrysanthem shengdi MOL004492 585 8.24 38.72 0.51 -1 0.6 0.3 17.5 axanthin 20- shengdi MOL011455 Hexadecano 418.6 1.91 32.7 -0.24 -0.4 0.7 0.29 104 ylingenol huanglian MOL001454 berberine 336.4 3.45 36.86 1.24 0.57 0.8 0.19 6.57 huanglian MOL013352 Obacunone 454.6 2.68 43.29 0.01 -0.4 0.8 0.31 -13 huanglian MOL002894 berberrubine 322.4 3.2 35.74 1.07 0.17 0.7 0.24 6.46 huanglian MOL002897 epiberberine 336.4 3.45 43.09 1.17 0.4 0.8 0.19 6.1 huanglian MOL002903 (R)-Canadine 339.4 3.4 55.37 1.04 0.57 0.8 0.2 6.41 huanglian MOL002904 Berlambine 351.4 2.49 36.68 0.97 0.17 0.8 0.28 7.33 Corchorosid huanglian MOL002907 404.6 1.34 105 -0.91 -1.3 0.8 0.29 6.68 e A_qt Magnogrand huanglian MOL000622 266.4 1.18 63.71 0.02 -0.2 0.2 0.3 3.17 iolide huanglian MOL000762 Palmidin A 510.5 4.52 35.36 -0.38 -1.5 0.7 0.39 33.2 huanglian MOL000785 palmatine 352.4 3.65 64.6 1.33 0.37 0.7 0.13 2.25 huanglian MOL000098 quercetin 302.3 1.5 46.43 0.05 -0.8 0.3 0.38 14.4 huanglian MOL001458 coptisine 320.3 3.25 30.67 1.21 0.32 0.9 0.26 9.33 huanglian MOL002668 Worenine
    [Show full text]
  • Pentosan Polysulfate Binds to STRO
    Wu et al. Stem Cell Research & Therapy (2017) 8:278 DOI 10.1186/s13287-017-0723-y RESEARCH Open Access Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis Jiehua Wu1,7, Susan Shimmon1,8, Sharon Paton2, Christopher Daly3,4,5, Tony Goldschlager3,4,5, Stan Gronthos6, Andrew C. W. Zannettino2 and Peter Ghosh1,5* Abstract Background: The pharmaceutical agent pentosan polysulfate (PPS) is known to induce proliferation and chondrogenesis of mesenchymal progenitor cells (MPCs) in vitro and in vivo. However, the mechanism(s) of action of PPS in mediating these effects remains unresolved. In the present report we address this issue by investigating the binding and uptake of PPS by MPCs and monitoring gene expression and proteoglycan biosynthesis before and after the cells had been exposed to limited concentrations of PPS and then re-established in culture in the absence of the drug (MPC priming). Methods: Immuno-selected STRO-1+ mesenchymal progenitor stem cells (MPCs) were prepared from human bone marrow aspirates and established in culture. The kinetics of uptake, shedding, and internalization of PPS by MPCs was determined by monitoring the concentration-dependent loss of PPS media concentrations using an enzyme-linked immunosorbent assay (ELISA) and the uptake of fluorescein isothiocyanate (FITC)-labelled PPS by MPCs. The proliferation of MPCs, following pre-incubation and removal of PPS (priming), was assessed using the Wst-8 assay 35 method, and proteoglycan synthesis was determined by the incorporation of SO4 into their sulphated glycosaminoglycans.
    [Show full text]
  • The KMT1A-GATA3-STAT3 Circuit Is a Novel Self-Renewal Signaling of Human Bladder Cancer Stem Cells Zhao Yang1, Luyun He2,3, Kais
    The KMT1A-GATA3-STAT3 circuit is a novel self-renewal signaling of human bladder cancer stem cells Zhao Yang1, Luyun He2,3, Kaisu Lin4, Yun Zhang1, Aihua Deng1, Yong Liang1, Chong Li2, 5, & Tingyi Wen1, 6, 1CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China 2Core Facility for Protein Research, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China 3CAS Key Laboratory of Infection and Immunity, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China 4Department of Oncology, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China 5Beijing Jianlan Institute of Medicine, Beijing 100190, China 6Savaid Medical School, University of Chinese Academy of Sciences, Beijing 100049, China Correspondence author: Tingyi Wen, e-mail: [email protected] Chong Li, e-mail: [email protected] Supplementary Figure S1. Isolation of human bladder cancer stem cells. BCMab1 and CD44 were used to isolate bladder cancer stem cells (BCSCs: BCMab1+CD44+) and bladder cancer non-stem cells (BCNSCs: BCMab1-CD44-) from EJ, samples #1 and #2 by flow cytometry. Supplementary Figure S2. Gene ontology analysis of downregulated genes of human BCSCs. (A) Pathway enrichment of 103 downregulated genes in BCSCs. (B) The seven downregulated genes in BCSCs participating in centromeric heterochromatin, mRNA-3’-UTR binding and translation regulator activity signaling pathways were validated by qRT-PCR. Data are presented as mean ± SD. P < 0.05; P < 0.01. Supplementary Figure S3. The expression of KMT1A is higher in human BC than that in peri-tumor tissues. (A) The expression of KMT1A was higher in BC samples than that in peri-tumors as assessed by immunohistochemistry, Scale bar = 50 m.
    [Show full text]
  • Genomic Organization of the Approximately 1.5 Mb Smith
    European Journal of Human Genetics (2001) 9, 892 ± 902 ã 2001 Nature Publishing Group All rights reserved 1018-4813/01 $15.00 www.nature.com/ejhg ARTICLE Genomic organisation of the ~1.5 Mb Smith-Magenis syndrome critical interval: Transcription map, genomic contig, and candidate gene analysis Rebecca E Lucas1, Christopher N Vlangos1, Parimal Das4, Pragna I Patel4 and Sarah H Elsea*,1,2,3 1Genetics Graduate Program, Michigan State University, East Lansing, Michigan, MI 48824, USA; 2Department of Zoology, Michigan State University, East Lansing, Michigan, MI 48824, USA; 3Department of Pediatrics and Human Development, Michigan State University, East Lansing, Michigan, MI 48824, USA; 4Department of Neurology, Baylor College of Medicine, Houston, Texas, TX 77030, USA Smith-Magenis syndrome (SMS) is a multiple congenital anomalies/mental retardation syndrome associated with an interstitial deletion of chromosome 17 involving band p11.2. SMS is hypothesised to be a contiguous gene syndrome in which the phenotype arises from the haploinsufficiency of multiple, functionally-unrelated genes in close physical proximity, although the true molecular basis of SMS is not yet known. In this study, we have generated the first overlapping and contiguous transcription map of the SMS critical interval, linking the proximal 17p11.2 region near the SMS-REPM and the distal region near D17S740 in a minimum tiling path of 16 BACs and two PACs. Additional clones provide greater coverage throughout the critical region. Not including the repetitive sequences that flank the critical interval, the map is comprised of 13 known genes, 14 ESTs, and six genomic markers, and is a synthesis of Southern hybridisation and polymerase chain reaction data from gene and marker localisation to BACs and PACs and database sequence analysis from the human genome project high-throughput draft sequence.
    [Show full text]
  • Loss of Supervillin Causes Myopathy with Myofibrillar Disorganization and Autophagic Vacuoles
    University of Massachusetts Medical School eScholarship@UMMS Open Access Articles Open Access Publications by UMMS Authors 2020-08-01 Loss of supervillin causes myopathy with myofibrillar disorganization and autophagic vacuoles Carola Hedberg-Oldfors University of Gothenburg Et al. Let us know how access to this document benefits ou.y Follow this and additional works at: https://escholarship.umassmed.edu/oapubs Part of the Amino Acids, Peptides, and Proteins Commons, Cardiovascular Diseases Commons, Cell Biology Commons, Cellular and Molecular Physiology Commons, Molecular and Cellular Neuroscience Commons, Musculoskeletal Diseases Commons, Musculoskeletal, Neural, and Ocular Physiology Commons, Nervous System Diseases Commons, and the Neurology Commons Repository Citation Hedberg-Oldfors C, Luna EJ, Oldfors A, Knopp C. (2020). Loss of supervillin causes myopathy with myofibrillar disorganization and autophagic vacuoles. Open Access Articles. https://doi.org/10.1093/ brain/awaa206. Retrieved from https://escholarship.umassmed.edu/oapubs/4320 Creative Commons License This work is licensed under a Creative Commons Attribution-Noncommercial 4.0 License This material is brought to you by eScholarship@UMMS. It has been accepted for inclusion in Open Access Articles by an authorized administrator of eScholarship@UMMS. For more information, please contact [email protected]. doi:10.1093/brain/awaa206 BRAIN 2020: 143; 2406–2420 | 2406 Loss of supervillin causes myopathy with Downloaded from https://academic.oup.com/brain/article/143/8/2406/5890852 by Medical Center Library user on 17 September 2020 myofibrillar disorganization and autophagic vacuoles Carola Hedberg-Oldfors,1,* Robert Meyer,2,* Kay Nolte,3,* Yassir Abdul Rahim,1 Christopher Lindberg,4 Kristjan Karason,5 Inger Johanne Thuestad,6 Kittichate Visuttijai,1 Mats Geijer,7,8 Matthias Begemann,2 Florian Kraft,2 Eva Lausberg,2 Lea Hitpass,9 Rebekka Go¨tzl,10 Elizabeth J.
    [Show full text]
  • Open Data for Differential Network Analysis in Glioma
    International Journal of Molecular Sciences Article Open Data for Differential Network Analysis in Glioma , Claire Jean-Quartier * y , Fleur Jeanquartier y and Andreas Holzinger Holzinger Group HCI-KDD, Institute for Medical Informatics, Statistics and Documentation, Medical University Graz, Auenbruggerplatz 2/V, 8036 Graz, Austria; [email protected] (F.J.); [email protected] (A.H.) * Correspondence: [email protected] These authors contributed equally to this work. y Received: 27 October 2019; Accepted: 3 January 2020; Published: 15 January 2020 Abstract: The complexity of cancer diseases demands bioinformatic techniques and translational research based on big data and personalized medicine. Open data enables researchers to accelerate cancer studies, save resources and foster collaboration. Several tools and programming approaches are available for analyzing data, including annotation, clustering, comparison and extrapolation, merging, enrichment, functional association and statistics. We exploit openly available data via cancer gene expression analysis, we apply refinement as well as enrichment analysis via gene ontology and conclude with graph-based visualization of involved protein interaction networks as a basis for signaling. The different databases allowed for the construction of huge networks or specified ones consisting of high-confidence interactions only. Several genes associated to glioma were isolated via a network analysis from top hub nodes as well as from an outlier analysis. The latter approach highlights a mitogen-activated protein kinase next to a member of histondeacetylases and a protein phosphatase as genes uncommonly associated with glioma. Cluster analysis from top hub nodes lists several identified glioma-associated gene products to function within protein complexes, including epidermal growth factors as well as cell cycle proteins or RAS proto-oncogenes.
    [Show full text]
  • Human Induced Pluripotent Stem Cell–Derived Podocytes Mature Into Vascularized Glomeruli Upon Experimental Transplantation
    BASIC RESEARCH www.jasn.org Human Induced Pluripotent Stem Cell–Derived Podocytes Mature into Vascularized Glomeruli upon Experimental Transplantation † Sazia Sharmin,* Atsuhiro Taguchi,* Yusuke Kaku,* Yasuhiro Yoshimura,* Tomoko Ohmori,* ‡ † ‡ Tetsushi Sakuma, Masashi Mukoyama, Takashi Yamamoto, Hidetake Kurihara,§ and | Ryuichi Nishinakamura* *Department of Kidney Development, Institute of Molecular Embryology and Genetics, and †Department of Nephrology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan; ‡Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Hiroshima, Japan; §Division of Anatomy, Juntendo University School of Medicine, Tokyo, Japan; and |Japan Science and Technology Agency, CREST, Kumamoto, Japan ABSTRACT Glomerular podocytes express proteins, such as nephrin, that constitute the slit diaphragm, thereby contributing to the filtration process in the kidney. Glomerular development has been analyzed mainly in mice, whereas analysis of human kidney development has been minimal because of limited access to embryonic kidneys. We previously reported the induction of three-dimensional primordial glomeruli from human induced pluripotent stem (iPS) cells. Here, using transcription activator–like effector nuclease-mediated homologous recombination, we generated human iPS cell lines that express green fluorescent protein (GFP) in the NPHS1 locus, which encodes nephrin, and we show that GFP expression facilitated accurate visualization of nephrin-positive podocyte formation in
    [Show full text]
  • Downloaded the Amino Acid Sequence and Used PSI-BLAST (Altschul Et Al
    bioRxiv preprint doi: https://doi.org/10.1101/2021.01.01.425057; this version posted January 3, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Homology between the flagellar export apparatus and ATP synthetase: evidence from synteny predating the Last Universal Common Ancestor Nicholas J. Matzke1, Angela Lin2, Micaella Stone1, Matthew A. B. Baker2,3* 1School of Biological Sciences, University of Auckland, Auckland 1010, New Zealand 2School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, Australia 3CSIRO Synthetic Biology Future Science Platform, Brisbane, Australia. *correspondence: [email protected] Abstract Evidence of homology between proteins in the ATP synthetase and the bacterial flagellar motor (BFM) has been accumulating since the 1980s. Specifically, the BFM’s Type 3 Secretion System (T3SS) export apparatus FliH, FliI, and FliJ are considered homologous to FO-b + F1-δ, F1-α/β, and F1-γ, and have similar structure and interactions. We review the discoveries that advanced the homology hypothesis and then conduct a further test by examining gene order in the two systems and their relatives. Conservation of gene order, or synteny, is often observed between closely related prokaryote species, but usually degrades with phylogenetic distance. As a result, observed conservation of synteny over vast phylogenetic distances can be evidence of shared ancestral coexpression, interaction, and function. We constructed a gene order dataset by examining the order of fliH, fliI, and fliJ genes across the phylogenetic breadth of flagellar and nonflagellar T3SS.
    [Show full text]