T-Cell Factor 1 Is a Gatekeeper for T-Cell Specification in Response to Notch Signaling

Total Page:16

File Type:pdf, Size:1020Kb

T-Cell Factor 1 Is a Gatekeeper for T-Cell Specification in Response to Notch Signaling T-cell factor 1 is a gatekeeper for T-cell specification in response to Notch signaling Kristine Germara,b, Marei Dosea, Tassos Konstantinoua, Jiangwen Zhangc, Hongfang Wangd, Camille Lobrye, Kelly L. Arnettf, Stephen C. Blacklowf, Iannis Aifantise, Jon C. Asterd, and Fotini Gounaria,1 aKnapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL 60637; bGraduate Program in Immunology, Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, MA 02111; cFaculty of Arts and Sciences, Research Computing, Harvard University, Cambridge, MA 02138; dDepartment of Pathology, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115; eHoward Hughes Medical Institute and Department of Pathology, NYU Cancer Institute, and Helen L. and Martin S. Kimmel Center for Stem Cell Biology, New York University School of Medicine, New York, NY 10016; and fDepartment of Cancer Biology, Dana-Farber Cancer Institute, and Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115 Edited* by Harvey Cantor, Dana-Farber Cancer Institute, Boston, MA, and approved October 19, 2011 (received for review June 27, 2011) Although transcriptional programs associated with T-cell specifica- function has not been linked with the up-regulation of T-lineage– tion and commitment have been described, the functional hierar- specific genes (16) or enhancement of T-cell development (17–19). chy and the roles of key regulators in structuring/orchestrating In this article, we establish that Tcf-1 is required in the earliest these programs remain unclear. Activation of Notch signaling in un- stages of T-cell specification. In the absence of Tcf-1, BM pro- committed precursors by the thymic stroma initiates the T-cell genitors entering the thymus fail to progress any further, and this differentiation program. One regulator first induced in these pre- effect is cell-intrinsic. The earliest defect detected in Tcf-1–de- fi cursors is the DNA-binding protein T-cell factor 1 (Tcf-1), a T-cell– cient thymocytes is the reduced expression of c-kit at the DN1 – fi specific mediator of Wnt signaling. However, the specific contri- stage of development. Tcf-1 de cient cells at this stage show increased apoptosis and have significantly reduced expression of bution of Tcf-1 to early T-cell development and the signals in- fi ducing it in these cells remain unclear. Here we assign functional genes involved in DNA metabolic processes, chromatin modi - significance to Tcf-1 as a gatekeeper of T-cell fate and show that cation, and response to damage compared with their WT coun- terparts. We further show that Notch binds the Tcf7 gene locus Tcf-1 is directly activated by Notch signals. Tcf-1 is required at the ∼ earliest phase of T-cell determinationforprogressionbeyondthe at a conserved element 31.5 kb upstream of the transcription start site, and ectopic expression of Notch1 leads to transcrip- early thymic progenitor stage. The global expression profile of tional up-regulation of Tcf-1. Thus, our data identify Tcf-1 as one Tcf-1–deficient progenitors indicates that basic processes of DNA of the earliest direct responders to Notch signaling in T-cell metabolism are down-regulated in its absence, and the blocked development. Tcf-1 is essential for thymic progenitors to proceed T-cell progenitors become abortive and die by apoptosis. Our through T-cell determination. data thus add an important functional relationship to the road- map of T-cell development. Results Tcf-1 Is Required for T-Cell Development Starting at the ETP Stage. cells are unique among hematopoietic cells because they need Earlier reports demonstrated that ablation of Tcf-1 causes an T a specialized organ, the thymus, to develop. Multipotent age-dependent degeneration of T-cell development (20). These progenitors (MPPs) migrating from the bone marrow (BM) studies showed that the DN1 population, thought at the time to through the blood stream seed the thymus (1, 2), whose micro- represent the earliest thymocyte subset, was abundantly present − − environment instructs them to lose stem cell properties and al- in Tcf-1 / mice, whereas the subsequent DN2, DN3, double- ternative developmental potentials and to differentiate along the positive, and single-positive populations were dramatically T-cell lineage (3). One important trigger for T-cell fate is the reduced. It is now understood that DN1 cells are highly het- activation of Notch signaling by Delta-like Notch ligands ex- erogeneous and can be subdivided into at least five subsets pressed on thymic stroma. Notch signaling induces the T-cell (DN1a–DN1e) (21). The c-kit–expressing DN1a and DN1b specification and commitment transcription programs (4). Com- subsets contain the most potent T-cell progenitors and are oth- mitment to the T-cell fate is endorsed as the cells progress from − − erwise known as ETPs (22). However, T cells can also be derived the early thymic progenitor (ETP; Lin CD44+c-kit+CD25 ), the from the atypical DN1c–DN1e subsets, although inefficiently most immature recognizable thymocyte, to the double-negative 3 (21). In light of these recent findings, we reanalyzed the earliest − − − − (DN3; Lin CD25+CD44 )stage.Aspartofthisprocess,co- thymic subpopulations in Tcf-1 / mice, including the classical − − − ordinated changes in the transcription profiles of these progeni- DN1 (Lin CD25 CD44+) (Fig. 1A), the ETP (Lin CD44+c- − tors gradually restrict their ability to become B, myeloid, dendritic, kit+CD25 ) (Fig. 1B), and the DN1a–DN1e subsets (Fig. 1C). − − and natural killer cells. Sustained Notch signaling is required Surprisingly, these analyses showed that Tcf-1 / thymi had during T-cell specification and commitment; however, Notch a previously unappreciated 100-fold reduction in absolute DN1 needs to collaborate with numerous other regulators in these numbers and a dramatic 300-fold reduction in ETPs compared − − − processes (5). Final commitment to the T-cell fate at the late DN2 with Tcf-1+/ or wt controls (Fig. 1D). Tcf-1 / DN1 cells had stage is marked by Bcl11b-mediated down-regulation of stem and reduced levels of surface c-kit (Fig. 1E), and an increased fraction progenitor cell transcriptional regulators (6–8). Thymus-like organs are present in all animals with adaptive immune systems, further emphasizing the specialized need for this organ (9, 10). Author contributions: K.G., M.D., T.K., H.W., and C.L. performed research; I.A., J.C.A., and Tcf-1 (product of the Tcf7 gene, referred to as Tcf-1 through- F.G. designed research; K.L.A., S.C.B., and J.C.A. contributed new reagents/analytic tools; out this article), is a T-lineage–specific, high-mobility group K.G., M.D., J.Z., H.W., and C.L. analyzed data; and K.G. and F.G. wrote the paper. (HMG) box-containing, DNA-binding protein. Tcf-1, like the The authors declare no conflict of interest. other T-cell factor/lymphoid enhancer-binding factor (Tcf/Lef) *This Direct Submission article had a prearranged editor. factors, mediates transcriptional activation when bound by Data deposition: The data reported in this paper have been deposited in the Gene Ex- β-catenin in response to Wnt signals or transcriptional repression pression Omnibus (GEO) database, www.ncbi.nlm.nih.gov/geo (accession no. GSE33513). when bound by Groucho (11). It has been well established that 1To whom correspondence should be addressed. E-mail: [email protected]. Tcf-1 is required in multiple stages of thymic and peripheral This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. T-cell development (12–15). However, Tcf/β-catenin gain of 1073/pnas.1110230108/-/DCSupplemental. 20060–20065 | PNAS | December 13, 2011 | vol. 108 | no. 50 www.pnas.org/cgi/doi/10.1073/pnas.1110230108 Downloaded by guest on September 29, 2021 − − Fig. 1. Early thymocyte development in Tcf-1 / mice. Pro- − − − files of gated Tcf-1 / and control Lin DN thymocytes are shown. (A) CD44 versus CD25 profiles. The DN1–DN4 subsets are indicated. Numbers indicate the frequencies of each subset. (BLeft) CD44 versus c-kit profiles show the electronic gating for pro-T cells. (Right) c-kit versus CD25 profiles of gated pro-T cells define the ETP, DN2a, and DN2b subsets as − − indicated. (C) Analysis of the DN1 subsets in Tcf-1 / and control mice. (Left) CD44 versus CD25 profiles show the gating of DN1 prothymocytes. (Right) CD24 versus c-kit profiles of gated DN1 cells define the DN1a (c-kit+CD24−), DN1b (c-kit+CD24+), DN1c (c-kitloCD24+), DN1d (c-kit−CD24+), − − and DN1e (c-kit CD24 ) subsets. (D) Bar histograms of cell − − − numbers in the indicated subsets of Tcf-1 / and Tcf-1+/ mice (n =4–6). (E) Histogram overlay of c-kit levels in gated DN1 thymocytes from the indicated mice. MFI, mean fluo- rescence intensity of c-kit. − − of cells expressed intermediate levels of CD25 (Fig. 1 A and C). In (Fig. 2E). Thus, Tcf-1 / progenitors are able to migrate agreement with earlier reports, DN2 cells and all subsequent through the bloodstream. thymic subsets were dramatically reduced. Thus, Tcf-1 is re- − − quired earlier than previously described: at, or before, thymic Tcf-1 / Progenitors Are Cell-Intrinsically Defective in T-Cell entry of uncommitted thymus-seeding progenitors. Although the Specification. The normal distribution of BM and blood progen- − − − − profile of early thymocytes in Tcf-1 / mice resembled that of itors compared with the reduced presence of Tcf-1 / ETPs fl fl fl Notch1-deficient, Notch2-haploinsufficient (MxCreN1 / N2 /+) suggests a functional inability to progress along the T-cell line- thymocytes (Fig. S1), no accumulation of B cells or macrophages age. To address this possibility, we examined the T-cell potential −/− was observed (Fig. S2), indicating that Tcf-1 may not be involved of hematopoietic progenitors from Tcf-1 mice in cocultures in the suppression of B-cell fate.
Recommended publications
  • Road Safety Investment Program in Romania - AA-010269
    CONSOLIDATED REPORT Framework agreement to support EIB advisory services (EIBAS) activities inside and outside EU-28 Lot 3: Transport Road Safety Investment Program in Romania - AA-010269 CONSULTING SAFEGE De Kleetlaan 5 B-1831 DIEGEM International Division SAFEGE SAS - SIÈGE SOCIAL Parc de l’Ile - 15/27 rue du Port 92022 NANTERRE CEDEX www.safege.com The authors take full responsibility for the contents of this report. The opinions expressed do not necessarily reflect the view of the Advisory Hub, nor the European Investment Bank, nor the European Commission Road Safety Investment Program in Romania - AA-010269 Framework agreement to support EIB advisory services (EIBAS) activities inside and outside EU-28 Lot 3: Transport Document quality information General information Author(s) Ilie Bricicaru, Kristiana Chakarova, Matthew Chamberlain, Loreta Robertina Gherman, Razvan Iulian Mazilu Project name RSIP in Romania – AA-010269 Document name Consolidated Report on Road Safety Investment Program in Romania Date April 05th, 2021 Reference Version 4 Addressee(s) Sent to: Name Organisation Sent on (date): Kevin CHEUNG, Per MATHIASEN, Teodora TATARU EIB Borislava GABROVSKA EIAH, EIB 05.04.2021 Cristian Andrei, Flavius PAVAL, Denyssa PPELIN CNAIR Copy to: Name Organisation Sent on (date): Joanna TALLEC, Spiros TRIANTAFILLOS SUEZ Consulting 05.04.2021 History of modifications Version Date Written by Approved & signed by: Ilie BRICICARU, Kristiana CHAKAROVA, Version 1 20.01.2021 Matthew CHAMBERLAIN Loreta Robertina GHERMAN Razvan Iulian MAZILU Ilie
    [Show full text]
  • Il Calendario Dei Divieti Di Circolazione Della Grecia E Della Spagna Non È Ancora Disponibile
    Driving restrictions, 2008 Austria 1. GENERAL DRIVING RESTRICTIONS Vehicles concerned Trucks with trailers, if the maximum authorised total weight of the motor vehicle or the trailer exceeds 3.5t; trucks, articulated vehicles and self-propelled industrial machines with an authorised total weight of more than 7.5t. Area Nationwide, with the exception of journeys made exclusively as part of a combined transport operation within a radius of 65km of the following transloading stations: Brennersee; Graz-Ostbahnhof; Salzburg-Hauptbahnhof; Wels-Verschiebebahnhof; Villach-Fürnitz; Wien-Südbahnhof; Wien-Nordwestbahnhof; Wörg; Hall in Tirol CCT; Bludenz CCT; Wolfurt CCT. Prohibition Saturdays from 15h00 to 24h00; Sundays and public holidays from 00h00 to 22h00 Public holidays 2008 1 January New Year’s Day 6 January Epiphany 24 March Easter Monday 1 May Labour Day; Ascension 12 May Whit Monday 22 May Corpus Christi 15 August Assumption 26 October National holiday 1 November All Saints’ Day 8 December Immaculate Conception 25 December Christmas Day 26 December Boxing Day Exceptions concerning trucks with trailers exceeding 3.5t · vehicles transporting milk; concerning vehicles with an authorised total weight of more than 7.5t · vehicles carrying meat or livestock for slaughter (but not the transport of heavy livestock on motorways), perishable foodstuffs (but not deep frozen goods), the supply of refreshments to tourist areas, urgent repairs to refrigeration plant, towing services (in all cases, according to § 46 StVO, it is obligatory to leave the motorway at the nearest exit), breakdown assistance vehicles, emergency vehicles, vehicles of a scheduled transport company (regular lines), and local trips on the two Saturdays preceding 24 December.
    [Show full text]
  • Raport Preliminar Asupra Master Planului Pe Termen Scurt Mediu Si Lung
    Raport preliminar asupra Master Planului pe termen scurt Mediu si Lung 19-08-2013 Preliminary Report on the Master Plan Short, Long and Medium Term 19-08- 2013 Transportation Guvernul României August 2013 Ministerul Transporturilor Master Planul General de Transport al României Versiune preliminară a Master Planului pe Termen Scurt, Mediu și Lung Elaborat de: ............................................................. Verificat de: Andrew Coates Simon Temple Frank Mohan Director Geoff Clarke Craig Bell Brian Vaughan Aprobat de: Martin Bright Director Error! Reference source not found.Raport Preliminar asupra Master Planului pe Termen Scurt, Mediu și Lung Rev No Comentarii Verificat de Aprobat de Data 1 Versiune preliminară SRT MJB 19/08/2013 Colmore Plaza, Colmore Circus Queensway, Birmingham, B4 6AT Telephone: 0121 262 1900 Website: http://www.aecom.com Job No: 60268467 Ref: Versiune preliminară Master Plan Data: August 2013 This document has been prepared by AECOM Limited for the sole use of our client (the “Client”) and in accordance with generally accepted consultancy principles, the budget for fees and the terms of reference agreed between AECOM Limited and the Client. Any information provided by third parties and referred to herein has not been checked or verified by AECOM Limited, unless otherwise expressly stated in the document. No third party may rely upon this document without the prior and express written agreement of AECOM Limited. 1 Cuprins 1 Scopul și obiectivele principale ale Master Planului ....................................................................................................
    [Show full text]
  • PROHIBICIONES DE CIRCULACION PARA VEHICULOS PESADOS EN EUROPA ( Desde El 01.01.2011 Hasta El 31.12.2011 )
    Madrid, 3 de Febrero 2011 ASTIC CIRCULAR Nº. 1.912 PROHIBICIONES DE CIRCULACION PARA VEHICULOS PESADOS EN EUROPA ( Desde el 01.01.2011 hasta el 31.12.2011 ) Ö EL INDICE de países figura en la última página (nº 58) A L E M A N I A Prohibición Para los vehículos de más de 7,5 t. de PMA, así como para camiones con remolque. Localización En toda la red de carreteras y autopistas. Período Domingos y días festivos de 00’00h. a 22’00h. Prohibición estival Localización Determinados tramos de autopistas y carreteras nacionales: - A1- de Cologne oeste vía Wuppertal, el cruce de Kamen y Münster hasta la salida a Cloppenburg, y de la entrada a Oyten hasta el cruce de Horst - A2- del cruce Oberhausen hasta el cruce Bad Oeynhausen - A3- del cruce Oberhausen hasta el cruce Cologne este, del cruce Mönchhof vía la intersección Francfort hasta la intersección Nürnberg - A4/E40- de la entrada a Herleshausen hasta el cruce Nossen - A5- del cruce Darmstadt vía Karlsruhe hasta el cruce Neuenburg - A6- de la entrada a Schwetzingen-Hockenheim hasta el cruce Nürnberg-sur - A7- de la entrada a Schleswig/Jagel hasta la salida Hambourg- Schnelsen-norte, del cruce Soltau-este al cruce Göttingen-nord; del cruce Schweinfurt/Werneck, el cruce Biebelried, el cruce Ulm/Elchingen, el cruce Allgäu hasta la frontera nacional en Füssen 2 - A8- del cruce Karlsruhe hasta la salida Munich-oeste y de la entrada a Munich- Ramersdorf hasta la salida Bad Reichenhall - A9/E51- circunvalación de Berlín (cruce Leipzig/cruce Potsdam) hasta la salida Munich-Schwabing - A10- circunvalación
    [Show full text]
  • RT520 LID Board
    130 Broadband Seismic Recorder RT520 Board Lid Interconnect -B01 130-01/-02 Refraction Technology™ RT520 Lid Interconnect 130-RT520-D 6/23/04 Refraction Technology, Inc. 2626 Lombardy Lane, Suite 105 Dallas, Texas 75220 USA Telephone: (214) 353-0609 FAX: (214) 353-9659 EMAIL: [email protected] FTP: ftp.reftek.com WWW: http://reftek.com This document is located at:m:\130\boards\RT520D\docs\RT520D.book Copyright 2004 Refraction Technology, Inc. All rights reserved. Printed in USA ii RT520 Lid Interconnect 130-RT520-D REF TEK Update Notification You can receive E-mail notification of updates by subscribing to one of the REF TEK 130 notification lists. To subscribe: 1 Send an E-mail to [email protected] with one of the following in the body of the message: “subscribe <listname> <your name> where <list name> is one of the entries in the following table and <your name> is replaced by your name. 2 For the REF TEK 130 Strong Motion (130-SM) product use rt_130_sm. DAS <listname> example 130 rt_130 subscribe rt_130 tom smith 130_ANSS rt_130_anss subscribe rt_130_anss john doe 130_SM rt_130_sm subscribe rt_130_sm mike smith 3 Your E-mail address is automatically extracted from the header of the E-Mail you send. To obtain updates for REF TEK software, firmware, and documents: [1] Login to our FTP site at: ftp.reftek.com\pub [2] User name: Anonymous [3] Password: Your E-mail Address Revision History: Rev Date Reason for change Affected Pages 0.1 7/05/03 Initial release All C 8/13/03 Created separate RT520C book with three B0 All versions (-B01, -B02, -B03) D 9/19/03 Improved filtering for all B0 versions All RT520 Lid Interconnect 130-RT520-D iii Notation Conventions The following notation conventions are used throughout Ref Tek documentation: Notation Description ASCII Indicates the entry conforms to the American Standard Code for Information Interchange definition of character (text) information.
    [Show full text]
  • Driving Restrictions, Goods Transport Germany 2019 Vehicles Concerned Trucks with a Total Permissible Weight of Over 7.5T, As We
    Driving Restrictions, Goods Transport Germany 2019 Vehicles concerned trucks with a total permissible weight of over 7.5t, as well as trucks with trailers in case of business-like/commercial or paid transportation of goods including related empty runs. Area throughout the road and motorway network Prohibition Sundays and public holidays from 00h00 to 22h00 Exceptions (applies also to the additional summer driving restrictions) 1. Combined rail/road goods transport from the shipper to the nearest loading railway station or from the nearest designated unloading railway station to the consignee up to a distance of 200km (no limitation on distance during the additional summer restrictions); also combined sea/road goods transport between the place of loading or unloading and a port situated within a radius of 150km maximum (delivery or loading). 2. Deliveries of fresh milk and other dairy produce, fresh meat and its fresh derivatives, fresh fish, live fish and their fresh derivatives, perishable foodstuffs (fruit and vegetables). 3. Transportation of animal by-products according to category 1, Art. 8 as well as category 2, Art. 9f of regulation (EG) No. 1069/2009. 4. Use of vehicles of recovery, towing and breakdown services in case of an accident or other emergencies. 5. Transport of living bees. 6. Empty vehicles, in connection with the transport operations mentioned under point 2-5. 7. Transport operations using vehicles subject to the Federal Law on the obligations of service; the relevant authorisation must be carried on board and produced for inspection on request. Also exempted from the prohibition are vehicles belonging to the police and federal border guard, fire brigades and emergency services, the federal armed services and allied troops.
    [Show full text]
  • Anexa La Ordinul Ministrului Transporturilor Nr
    Anexa la Ordinul Ministrului Transporturilor nr. 1154/2019 Sectoarele de drumuri principale din administrarea CNAIR SA pentru care au fost intocmite harti strategice de zgomot in anul 2017 si datele privind expunerea la zgomotul generat de traficul rutier pe aceste sectoare Limite sector(km) Limite sector (STEREO70) Estimare numar persoane expuse Indicativ Lungime Nr. Crt de la la Limite sector drum sector Lzsn(dB) Ln (dB) de la la Xstart Ystart Xend Yend 55-60 60-65 65-70 70-75 >75 45-50 50 - 55 55 - 60 60 - 65 65 - 70 > 70 1 A1 10.600 30.510 577564.1443 326692.6011 558416.5221 329429.8287 19.910 580 489 254 249 144 753 507 359 198 196 43 M.Bucuresti - DJ401A 2 A1 30.510 49.208 558416.5221 329429.8287 542229.5877 337267.9619 18.698 159 85 31 9 0 188 119 40 18 5 0 DJ401A - DJ711A 3 A1 49.208 70.460 542229.5877 337267.9619 526962.8947 350829.8984 21.252 68 43 46 48 8 103 23 60 49 18 0 DJ711A - DN61 4 A1 70.460 94.393 526962.8947 350829.8984 506579.5499 361785.1586 23.933 89 42 33 27 2 112 60 39 19 15 0 DN61 - DJ703B 5 A1 94.393 106.500 506579.5499 361785.1586 496677.6883 367981.2796 12.107 141 72 54 10 0 144 101 62 26 0 0 DJ703B - DN65B 6 A1 106.500 120.100 496677.6883 367981.2796 487285.4852 377468.0906 13.600 1278 371 98 20 4 2362 488 191 35 5 0 A1 - DN7 7 A1 237.432 254.240 437958.4521 472395.1111 427976.6921 480103.6184 16.808 728 165 13 0 0 1197 261 65 0 0 0 DN14 - DN7H (DJ106B) 8 A1 254.240 270.345 427976.6921 480103.6184 414523.3048 479569.1638 16.105 64 14 13 0 0 85 28 16 2 0 0 DN7H (DJ106B) - DN1 9 A1 292.455 303.600 400937.6833 491830.5315 391041.4987 494901.1517 11.145 12 5 0 0 0 19 7 1 0 0 0 N.
    [Show full text]
  • Guvernul României
    GUVERNUL ROMÂNIEI HOTǍRÂRE pentru modificarea şi completarea Hotǎrârii Guvernului nr. 321/2005 privind evaluarea şi gestionarea zgomotului ambiant, republicată În temeiul art. 108 din Constituţia României, republicată, Guvernul României adoptă prezenta hotărâre. Articolul I – Hotărârea Guvernului nr. 321/2005 privind evaluarea şi gestionarea zgomotului ambiant, republicată în Monitorul Oficial al României, Partea I, nr. 19 din 10 ianuarie 2008, se modifică şi se completează după cum urmează: 1. La articolul 1, partea introductivă a alineatului (1) se modifică şi va avea următorul cuprins: “(1)Prezenta hotărâre abordează unitar la nivel naţional, evitarea, prevenirea sau reducerea efectelor dăunătoare, inclusiv a disconfortului, provocate de expunerea populaţiei la zgomotul ambiant, prin implementarea progresivă a următoarelor măsuri:” 2. La articolul 4, alineatul (1) se modifică şi va avea următorul cuprins: “(1) Autorităţile administraţiei publice locale realizează cartarea zgomotului şi elaborează hărţile strategice de zgomot şi planurile de acţiune potrivit prevederilor prezentei hotărâri, pentru aglomerările, drumurile judeţene şi aeroporturile aflate în administrarea lor, cu respectarea termenelor prevăzute la alin. (7) şi (8) şi implementează măsurile de reducere şi gestionare a zgomotului cu respectarea termenelor care se menţionează în acest sens în planurile de acţiune. 3. La articolul 4, alineatul (5) şi alineatul (6) se modifică şi vor avea următorul cuprins: “(5) Toate autorităţile administraţiei publice şi toţi operatorii
    [Show full text]
  • You Drive, We Care
    You drive, we care. RO - Diesel & Services Rumänien / România / Romania PLZ sortiert Sorted by ZIP code » For help, call me! DKV ASSIST - 24h International Free Call* 00800 365 24 365 In case of difficulties concerning the number 00800 please dial the relevant emergency number of the country: Bei unerwarteten Schwierigkeiten mit der Rufnummer 00800, wählen Sie bitte die Notrufnummer des Landes: Andorra / Andorra Latvia / Lettland » +34 934 6311 81 » +370 5249 1109 Austria / Österreich Liechtenstein / Liechtenstein » +43 362 2723 03 » +39 047 2275 160 Belarus / Weißrussland Lithuania / Litauen » 8 820 0071 0365 (national) » +370 5249 1109 » +7 495 1815 306 Luxembourg / Luxemburg Belgium / Belgien » +32 112 5221 1 » +32 112 5221 1 North Macedonia / Nordmazedonien Bosnia-Herzegovina / Bosnien-Herzegowina » +386 2616 5826 » +386 2616 5826 Moldova / Moldawien Bulgaria / Bulgarien » +386 2616 5826 » +359 2804 3805 Montenegro / Montenegro Croatia / Kroatien » +386 2616 5826 » +386 2616 5826 Netherlands / Niederlande Czech Republic / Tschechische Republik » +49 221 8277 9234 » +420 2215 8665 5 Norway / Norwegen Denmark / Dänemark » +47 221 0170 0 » +45 757 2774 0 Poland / Polen Estonia / Estland » +48 618 3198 82 » +370 5249 1109 Portugal / Portugal Finland / Finnland » +34 934 6311 81 » +358 9622 2631 Romania / Rumänien France / Frankreich » +40 264 2079 24 » +33 130 5256 91 Russia / Russland Germany / Deutschland » 8 800 7070 365 (national) » +49 221 8277 564 » +7 495 1815 306 Great Britain / Großbritannien Serbia / Serbien » 0 800 1975 520
    [Show full text]
  • GUVERNUL ROMÂNIEI H O T Ă R Â R E Pentru Modificarea Și
    GUVERNUL ROMÂNIEI H O T Ă R Â R E pentru modificarea și completarea Hotărârii Guvernului nr. 321/2005 privind evaluarea și gestionarea zgomotului ambiant În temeiul art. 108 din Constituția României, republicată, Guvernul României adoptă prezenta hotărâre: ART.I. Hotărârea Guvernului nr.321/2005 privind evaluarea şi gestionarea zgomotului ambiant, republicată în Monitorul Oficial al României, Partea I, nr. 19 din 10 ianuarie 2008, cu modificările și completările ulterioare, se modifică şi se completează după cum urmează: 1. La articolul 4, alineatul (3) se modifică și va avea următorul cuprins: “(3) Unităţile aflate sub autoritatea autorităţii publice centrale pentru transporturi, care au în administrare infrastructuri rutiere, feroviare, aeroportuare şi portuare, realizează cartarea zgomotului şi elaborează hărţile strategice de zgomot şi planurile de acţiune, potrivit prevederilor prezentei hotărâri, pentru drumurile principale din interiorul si din exteriorul aglomerărilor, căile ferate principale din interiorul și din exteriorul aglomerărilor și căile ferate din interiorul aglomerărilor, aeroporturile mari, aeroporturile civile urbane din interiorul aglomerărilor şi porturile din interiorul aglomerărilor, aflate în administrarea acestora, cu respectarea prevederilor alin. (9) şi (10), şi implementează măsurile de reducere şi gestionare a zgomotului cu respectarea termenelor care se menţionează în acest sens în planurile de acţiune”. 2. La articolul 4, după alineatul (15) se introduce un nou alineat, alin.(151) care va avea următorul cuprins: “(151) Aglomerările, drumurile principale, căile ferate principale, aeroporturile civile urbane şi aeroporturile mari, precum şi porturile maritime şi/sau fluviale pentru care trebuie realizate hărţile strategice de zgomot şi datele aferente acestora până la data de 30 iunie 2017 şi planurile de acţiune aferente până la data de 18 iulie 2018, sunt prevăzute în anexa nr.
    [Show full text]
  • Länderinformationen Rumänien - Besondere Vorschriften Nur Ein Teil Des Rumänischen Straßennetzes Ist Für Lkw Mit Hohen Achslasten Geeignet
    Länderinformationen Länderinformationen Rumänien - Besondere Vorschriften Nur ein Teil des rumänischen Straßennetzes ist für Lkw mit hohen Achslasten geeignet. In vielen Fällen kann deshalb nicht auf dem direkten Weg zu den Kunden gefahren werden. Bei Überschreitung der höchstzulässigen Abmessungen und Gewichte werden in Rumänien hohe Gebühren erhoben (siehe "Höchstzulässige Abmessungen und Gewichte"). Eine Warmweste muss im Fahrzeug mitgeführt werden. Rumänien - Bilaterale Verkehre Ab 1. Januar 2007: Es gelten die Bestimmungen für die Euro-Lizenz (siehe EG-Marktzugangsverordnung 1072/2009). Rumänien - Dreiländerverkehre Mit Rumänien wurde eine Vereinbarung getroffen, wonach die Gemeinschaftslizenz als bilaterale Fahrtgenehmigung für den Dreiländerverkehr mit Durchfahren des Heimatlandes auf dem verkehrsüblichen Weg anerkannt wird. Rumänien - Fährverbindungen Donau - Fährgebühren nach Bulgarien - Donaufähre zwischen Bechet und Orjahovo (südlich von Craiova) Gebühren für Einzelfahrzeuge und Fahrzeugkombinationen: Einzelfahrt* 42,00 US-$ / 35,00 - Lieferwagen 55,00 US-$ / 45,00 - Lkw - 95,00 US-$ / 80,00 Fahrzeugkombinationen - Fahrer 4,00 US-$ / 3,00 http://intranet.bgl-ev.de/infos/land_info/land.asp (1 von 20)02.08.2012 12:14:40 Länderinformationen *) Die Tarife sind in US-$ und festgelegt und sind nicht an die aktuelle Wechselkursrate gebunden. - Donaufähre zwischen Giurgiu und Ruse (südlich von Bukarest) Gebühren für Einzelfahrzeuge und Fahrzeugkombinationen: Einzelfahrt 35,00 US-$ / 35,00 - Einzelfahrzeuge - 40,00 US-$ / 40,00 Fahrzeugkombinationen - Fahrer 1,00 US-$ - Donaufähre zwischen Calafat und Vidin (südwestlich von Craiova) Gebühren für Einzelfahrzeuge und Fahrzeugkombinationen: Einzelfahrt* 30,00 US-$ / 35,00 - Lieferwagen 40,00 US-$ / 45,00 - Lkw - 70,00 US-$ / 80,00 Fahrzeugkombinationen - Fahrer 3,00 US-$ / 3,00 *) Die Tarife sind in US-$ und festgelegt und sind nicht an die aktuelle Wechselkursrate gebunden.
    [Show full text]
  • Visualization of the Earliest Steps of Gammadelta T Cell Development in the Adult Thymus
    Visualization of the earliest steps of gammadelta T cell development in the adult thymus. Prinz, I., Sansoni, A., Kissenpfennig, A., Ardouin, L., Malissen, M., & Malissen, B. (2006). Visualization of the earliest steps of gammadelta T cell development in the adult thymus. Nature Immunology, 7(9)(9), 995-1003. https://doi.org/10.1038/ni1371 Published in: Nature Immunology Document Version: Early version, also known as pre-print Queen's University Belfast - Research Portal: Link to publication record in Queen's University Belfast Research Portal General rights Copyright for the publications made accessible via the Queen's University Belfast Research Portal is retained by the author(s) and / or other copyright owners and it is a condition of accessing these publications that users recognise and abide by the legal requirements associated with these rights. Take down policy The Research Portal is Queen's institutional repository that provides access to Queen's research output. Every effort has been made to ensure that content in the Research Portal does not infringe any person's rights, or applicable UK laws. If you discover content in the Research Portal that you believe breaches copyright or violates any law, please contact [email protected]. Download date:28. Sep. 2021 ARTICLES Visualization of the earliest steps of cd T cell development in the adult thymus Immo Prinz1, Amandine Sansoni1, Adrien Kissenpfennig1,2, Laurence Ardouin1, Marie Malissen1 & Bernard Malissen1 The checkpoint in cd cell development that controls successful T cell receptor (TCR) gene rearrangements remains poorly characterized. Using mice expressing a reporter gene ‘knocked into’ the Tcrd constant region gene, we have characterized many of the events that mark the life of early cd cells in the adult thymus.
    [Show full text]