NS-1 and NS-2 proteins may act synergistically in the cytopathogenicity of parvovirus MVMp

Brandenburger, Anne-Nicole; Legendre, Daniel; Avalosse, Bernard; Rommelaere, Jean

doi:10.1016/0042-6822(90)90110-d

Cited by 89 publications

(69 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…18,19 The NS proteins, moreover, exert cytotoxic activity in transformed cells. 20,21 These various observations led to the idea that one might achieve tumorselective expression of a foreign gene by means of a parvoviral vector obtained by replacing the capsid protein genes with the foreign gene. 10 Vectors based on MVMp were constructed, expressing 46 The resulting recombinant virus stocks were purified and concentrated as described.…”

mentioning

confidence: 99%

Tumor-selective gene transduction and cell killing with an oncotropic autonomous parvovirus-based vector

Dupont¹,

Avalosse²,

Karim³

et al. 2000

Gene Ther

View full text Add to dashboard Cite

A recombinant MVMp of the fibrotropic strain of minute virus of mice (MVMp) expressing the chloramphenicol acetyltransferase reporter gene was used to infect a series of biologically relevant cultured cells, normal or tumor-derived, including normal melanocytes versus melanoma cells, normal mammary epithelial cells versus breast adenocarcinoma cells, and normal neurons or astrocytes versus glioma cells. As a reference cell system we used normal human fibroblasts versus the SV40-transformed fibroblast cell line NB324K. After infection, we observed good expression of the reporter gene in the different tumor cell types, but only poor expression if any in the corresponding normal cells. We also constructed a recombinant MVMp expressing the green fluorescent protein reporter gene and assessed by flow cytometry the efficiency of gene transduction into the different target cells. At a multiplicity of infection of 30, we observed

show abstract

mentioning

confidence: 99%

Tumor-selective gene transduction and cell killing with an oncotropic autonomous parvovirus-based vector

Dupont¹,

Avalosse²,

Karim³

et al. 2000

Gene Ther

View full text Add to dashboard Cite

show abstract

“…FRMCEJ clones were obtained by transfection of FRMCI4 cells with plasmid pSVneoEJ, which carries the c-Ha-ras-1 oncogene from the human bladder carcinoma cell line EJ (McKay et al, 1986), followed by the selection of geneticin (0-5 mg/ml)-resistant colonies. Transfections were carried out by the calcium phosphate precipitation technique described by Brandenburger et al (1990).…”

Section: Methodsmentioning

confidence: 99%

“…The amplification of input MVMp D N A was measured by hybridization at 30 h after infection with a multiplicity of 1 p.f.u./cell. As shown in Table 1 The NS-1 protein of autonomous parvoviruses is a multifunctional phosphoprotein, the expression of which has been associated with cell killing (Rhode, 1987;Osawa et al, 1988;Brandenburger et al, 1990;CailletFauquet et al, 1990). Therefore it was of interest to determine whether the varying sensitivity of the transformants tested to MVMp infection was related to the differential expression of this viral polypeptide.…”

Section: Oncogene-induced Modulation Of Mvmp Replicationmentioning

confidence: 99%

“…This oncotropism is consistent with the fact that a number of neoplastic or transformed cells cultivated in vitro are permissive to the lytic parvovirus life cycle, whereas 0001-0911 © 1992 SGM normal cells generally resist virus attack. This sensitization of transformed cells correlates with a stimulation of parvoviral gene expression, suggesting that some viral proteins may be cytotoxic (Rhode, 1987;Osawa et al, 1988;Cornelis et al, 1988b;van Hille et al, 1989;Brandenburger et al, 1990). It has been demonstrated recently that the intracellular accumulation of MVMencoded regulatory non-structural (NS) proteins is toxic for neoplastic human cells (Caillet-Fauquet et al, 1990).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Cooperation of oncogenes in cell transformation and sensitization to killing by the parvovirus minute virus of mice

Legrand

Mousset

Salomé

et al. 1992

Journal of General Virology

Self Cite

View full text Add to dashboard Cite

“…NS -1 is a pleotropic, mainly nuclear phosphoprotein. 3 It mediates the toxicity of MVM in transformed cells 4,5 and is involved in viral DNA replication and promoter regulation. 6 -8 NS -1 transactivates, for instance, the promoter of capsid proteins, VP -1 and VP -2.…”

mentioning

confidence: 99%

Construction and production of oncotropic vectors, derived from MVM(p), that share reduced sequence homology with helper plasmids

2002

View full text Add to dashboard Cite

The production of currently available vectors derived from autonomous parvoviruses requires the expression of capsid proteins in trans, from helper sequences. Cotransfection of a helper plasmid always generates significant amounts of replication -competent virus ( RCV ) that can be reduced by the integration of helper sequences into a packaging cell line. Although stocks of minute virus of mice ( MVM ) -based vectors with no detectable RCV could be produced by transfection into packaging cells; the latter appear after one or two rounds of replication, precluding further amplification of the vector stock. Indeed, once RCVs become detectable, they are efficiently amplified and rapidly take over the culture. Theoretically RCV -free vector stocks could be produced if all homology between vector and helper DNA is eliminated, thus preventing homologous recombination. We constructed new vectors based on the structure of spontaneously occurring defective particles of MVM. Based on published observations related to the size of vectors and the sequence of the viral origin of replication, these vectors were modified by the insertion of foreign DNA sequences downstream of the transgene and by the introduction of a consensus NS -1 nick site near the origin of replication to optimize their production. In one of the vectors the inserted fragment of mouse genomic DNA had a synergistic effect with the modified origin of replication in increasing vector production. Cancer Gene Therapy ( 2002 ) 9, 762 -770 doi:10.1038/sj.cgt.7700496Keywords: parvovirus; replication -competent virus; minute virus of mice; 2nd generation vectors T he genome of parvovirus minute virus of mice prototype strain [MVM( p )] is well characterized; it has been cloned and entirely sequenced. 1,2 Nevertheless, some aspects of the regulation of its gene expression and of particle production are still not well understood. The 5 -kb singlestranded, linear genome of MVM( p) encodes two sets of proteins from overlapping genes. The nonstructural proteins, NS -1 and NS -2, are expressed under control of the P4 promoter. NS -1 is a pleotropic, mainly nuclear phosphoprotein. 3 It mediates the toxicity of MVM in transformed cells 4,5 and is involved in viral DNA replication and promoter regulation. 6 -8 NS -1 transactivates, for instance, the promoter of capsid proteins, VP -1 and VP -2. 9,10 The different functions of NS -1 are modulated by differential phosphorylation in the course of the viral infectious cycle. 11,12 Replication of H -1 and MVM has been shown to take place in nuclear structures called PAR bodies (H -1 ) or APAR bodies (MVM) (autonomous parvovirus -associated replication bodies ) that are formed after infection. 13,14 Two adjacent elements, A and B, have been identified at the right side of the genome as being necessary in cis for efficient replication. These internal replication sequences ( IRS ) partially overlap the 3 0 extremity of capsid genes. They bind as yet nonidentified cellular protein complexes called MVM replication factors ( MRF ). 15 ...

show abstract

NS-1 and NS-2 proteins may act synergistically in the cytopathogenicity of parvovirus MVMp

Cited by 89 publications

References 28 publications

Tumor-selective gene transduction and cell killing with an oncotropic autonomous parvovirus-based vector

Tumor-selective gene transduction and cell killing with an oncotropic autonomous parvovirus-based vector

Cooperation of oncogenes in cell transformation and sensitization to killing by the parvovirus minute virus of mice

Construction and production of oncotropic vectors, derived from MVM(p), that share reduced sequence homology with helper plasmids

Contact Info

Product

Resources

About