Nervous and nonnervous cell transduction by recombinant adenoviruses that inducibly express the human PrP

被引:6
作者
Arrabal, S
Touchard, M
Mouthon, F
Klonjkowski, B
Deslys, JP
Dormont, D
Eloit, M [1 ]
机构
[1] Ecole Natl Vet Alfort, URA INRA, F-94704 Maisons Alfort, France
[2] CEA, Serv Neurovirol, DSV, DRM,CRSSA,EPHE, F-92265 Fontenay Aux Roses, France
关键词
D O I
10.1006/bbrc.2001.5208
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The study of the prion protein (PrP) physiological functions or its specific role in transmissible spongiform encephalopathies (TSE) requires new tools, particularly those able to induce PrP overexpression in a large range of cells, in vivo as well as in vitro. Here we describe the construction of two recombinant adenoviruses encoding the human PrP either with a valine at position 129 (AdTRVal) or a methionine (AdTRMet). Both genes were put under the control of the tetracycline-responsive promoter, allowing tight regulation of PrP expression. AdTRVal and AdTRMet induced high expression of the human PrP in CHO-KI cells and in organotypic brain slices in culture. The proteins expressed from these viruses exhibited a glycosylphosphatidyl inositol (GPI) anchor, proper glycosylation and sensitivity to proteinase K digestion. AdTRVal and AdTRMet will allow future studies on the human PrP and on the role of the codon 129 polyphormism in human TSE. (C) 2001 Academic Press.
引用
收藏
页码:623 / 632
页数:10
相关论文
共 29 条
[1]   Efficacy of replication-defective adenovirus-vectored vaccines: Protection following intramuscular injection is linked to promoter efficiency in muscle representative cells [J].
Ambriovic, A ;
Adam, M ;
Monteil, M ;
Paulin, D ;
Eloit, M .
VIROLOGY, 1997, 238 (02) :327-335
[2]   Comparative interleukin (IL)-2 interferon (IFN)-gamma and IL-4/IL-10 responses during acute infection of macaques inoculated with attenuated nef-truncated or pathogenic SIVmac251 virus [J].
Benveniste, O ;
Vaslin, B ;
LeGrand, R ;
Cheret, A ;
Matheux, F ;
Theodoro, F ;
Cranage, MP ;
Dormont, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (08) :3658-3663
[3]   Prion protein expression in Chinese hamster ovary cells using a glutamine synthetase selection and amplification system [J].
Blochberger, TC ;
Cooper, C ;
Peretz, D ;
Tatzelt, J ;
Griffith, OH ;
Baldwin, MA ;
Prusiner, SB .
PROTEIN ENGINEERING, 1997, 10 (12) :1465-1473
[4]   Normal prion protein has an activity like that of superoxide dismutase [J].
Brown, DR ;
Wong, BS ;
Hafiz, F ;
Clive, C ;
Haswell, SJ ;
Jones, IM .
BIOCHEMICAL JOURNAL, 1999, 344 :1-5
[5]   NORMAL DEVELOPMENT AND BEHAVIOR OF MICE LACKING THE NEURONAL CELL-SURFACE PRP PROTEIN [J].
BUELER, H ;
FISCHER, M ;
LANG, Y ;
BLUETHMANN, H ;
LIPP, HP ;
DEARMOND, SJ ;
PRUSINER, SB ;
AGUET, M ;
WEISSMANN, C .
NATURE, 1992, 356 (6370) :577-582
[6]   PRION PROTEIN IS NECESSARY FOR NORMAL SYNAPTIC FUNCTION [J].
COLLINGE, J ;
WHITTINGTON, MA ;
SIDLE, KCL ;
SMITH, CJ ;
PALMER, MS ;
CLARKE, AR ;
JEFFERYS, JGR .
NATURE, 1994, 370 (6487) :295-297
[7]   New insight into abnormal prion protein using monoclonal antibodies [J].
Demart, S ;
Fournier, JG ;
Creminon, C ;
Frobert, Y ;
Lamoury, F ;
Marce, D ;
Lasmézas, C ;
Dormont, D ;
Grassi, J ;
Deslys, JP .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 265 (03) :652-657
[8]   TIGHT CONTROL OF GENE-EXPRESSION IN MAMMALIAN-CELLS BY TETRACYCLINE-RESPONSIVE PROMOTERS [J].
GOSSEN, M ;
BUJARD, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (12) :5547-5551
[9]   CONTROL OF GENE ACTIVITY IN HIGHER EUKARYOTIC CELLS BY PROKARYOTIC REGULATORY ELEMENTS [J].
GOSSEN, M ;
BONIN, AL ;
BUJARD, H .
TRENDS IN BIOCHEMICAL SCIENCES, 1993, 18 (12) :471-475
[10]   NEW TECHNIQUE FOR ASSAY OF INFECTIVITY OF HUMAN ADENOVIRUS 5 DNA [J].
GRAHAM, FL ;
VANDEREB, AJ .
VIROLOGY, 1973, 52 (02) :456-467