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J C D'Halluin

Publications and source records attributed to J C D'Halluin.

At least 37 records · Page 2Linked to original sources
PubMed1988

The role of the two E1a mRNA products of subgroup B adenoviruses in the regulation of early promoters of subgroup C adenoviruses.

HeLa cells were co-transfected with recombinant plasmids carrying adenovirus (Ad)2 or Ad3 E1a promoters fused to the chloramphenicol acetyl transferase gene (cat), and a plasmid encoding the Ad3 E1a promoter. Whereas no stimulating effect was observed on the Ad3 E1a promoter, the Ad2 promoter was inhibited. To determine which of the E1a gene products of Ad3 was responsible for the repressive effect, plasmids were constructed in which only the 13S or 12S mRNA product of Ad3 was expressed. Both the 12S and 13S mRNA products of Ad3 E1a were found to depress the transcription from the Ad2 E1a promoter. Each Ad3 E1a gene product was able to stimulate transcription from the Ad5 E2a early promoter in a manner similar to that of the Ad2 E1a gene products. In the case of the Ad5 E3 promoter, neither of the Ad3 E1a gene products stimulated transcription, but an inhibition was observed. These results suggest that both mRNA products of the Ad3 E1a region inhibit transcription at the TATA box transcription complex.

Adenovirus Early Proteins↗
PubMed1988

Differences in the organization of adenovirus E1A promoters are not important for their full activity.

The 5'-E1A control regions of adenovirus (Ad) types 2, 3 and 12 were cloned upstream from the cat gene. Each serotype was characterized by unique organization of the 5'-E1A non-coding region. All three 5'-non-coding E1A regions were able to stimulate the cat gene transcription in HeLa cells. Hybrid plasmids between Ad2/Ad3 and Ad2/Ad12 revealed the same level of CAT activity as the native plasmids but the optimum activity of promoter was achieved only when its organization was not modified. These observations suggest that various nuclear factors take part in the activation process of each promoter.

Adenoviridae↗
PubMed1987

Expression of protein IIIa of human adenovirus type 2 in Escherichia coli.

We have constructed a plasmid encoding the protein IIIa gene of human adenovirus type 2. The gene was expressed under the control of the hybrid tac (trp-lac) promoter; the protein was synthesized at levels up to 5% of newly synthesized protein after IPTG induction. The protein IIIa produced in Escherichia coli has an apparent Mr on sodium dodecyl sulfate-polyacrylamide gels of 67 kDa, and was revealed with anti-adenovirus serum in Western blotting. The protein IIIa produced in bacteria was phosphorylated in the presence of [gamma-32P]ATP.

Adenoviruses, Human↗
PubMed1986

Expression of the chloramphenicol acetyl transferase gene in human cells under the control of early adenovirus subgroup C promoters: effect of E1A gene products from other subgroups on gene expression.

A hierarchy of dominance has been observed in HeLa cells co-infected with two serotypes of adenovirus belonging to different subgroups. DNA replication and late protein synthesis of one serotype are inhibited by those of the other. The degree of inhibitory effect has the following decreasing order: adenovirus type 3 (Ad3) and Ad7 (subgroup B), Ad9 (D), Ad4 (E), Ad12 (A), Ad2 and Ad5 (C) [Delsert and D'Halluin, Virus Res. 1 (1984) 365-380]. HeLa cells were first transfected with recombinant plasmids carrying Ad5 E2A or E3 promoters fused to the chloramphenicol acetyl transferase gene (cat), and then infected with human Ad belonging to different subgroups. All the serotypes tested were found to be able to stimulate both E2A and E3 promoters. When HeLa cells were co-transfected with either of the previous plasmids, plus a second plasmid carrying the Ad3 E1A region, the same stimulatory effect was observed. However, an inhibitory effect on Ad5 E2A and E3 promoters seemed to occur when both Ad2 E1A (subgroup C) and Ad3 E1A (subgroup B) genes were present together. To determine which one of the early products was responsible for the observed repression effect, and to assign the target on the genome of subgroup C Ad, a plasmid was constructed in which the sequences at the 5' end of the Ad2 E1A region were fused to the structural sequences of the cat gene. In HeLa cells transfected with this plasmid, CAT activity was significantly increased after co-transfection with a plasmid carrying the Ad2 E1A region, but decreased with a plasmid carrying the Ad3 E1A region.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetyltransferases↗
PubMed1986

Human adenovirus 2 temperature-sensitive mutant 112 contains three mutations in the protein IIIa gene.

The temperature-sensitive (ts) mutant 112 of human adenovirus 2 is defective in the late stage of virus maturation. The region of functional mutation has been localised by marker rescue. It was observed that the ts mutation can be rescued by the left-hand part of the wild-type gene (nucleotides 12,301-12,891). By nucleotide sequencing, two mutations, both C to T (at position 12,386 and 12,741), were found in this region. The first one, in the glycine 20 codon, is silent, whereas the second changes alanine 145 to valine. A third mutation, which changed C to A (nucleotide 13,613), was identified in the right-hand part of the gene, resulting in the replacement of alanine-436 by threonine.

Adenoviruses, Human↗
PubMed1985

An adenovirus cytocidal function related to the control of a cellular pH 4 endonuclease activity.

An adenovirus (Ad) interserotypic recombinant (H2cyt141) between temperature-sensitive mutant H2ts111 of Ad2 and deletion mutant H5dl313 of Ad5 was isolated and characterized. It was phenotypically ts+, dl+, hr+ and formed large plaques (or cytocidal: cyt). It contained the right 89% of Ad5 DNA and the leftmost 11% of Ad2 DNA. Genetic recombination data suggested the cytocidal mutation lay in the transforming region E1B, confirming sequence analysis. The cytocidal effect resulted in part from the breakdown of cellular DNA. Host cell and virus DNA breakdown induced by H2cyt141 appeared cell-dependent: it occurred in HeLa, KB or BHK-21 cells, but not in CV1 or 293 cells. In human cells the cyt effect was recessive and adenovirus DNA degradation was prevented by co-infection with adenovirus wild-type (H2WT), other adenovirus serotypes or simian virus 40 (SV40). In simian cells, H2cyt141 did not inhibit SV40 DNA replication, unlike H2WT. The amount of H2cyt141 DNA integrated in human cell DNA at early stages of the lytic cycle was found to be significantly lower than for H2WT. Novobiocin inhibited viral DNA breakdown in human cells. Cellular DNA extracted from H2cyt141-infected cells exhibited a repeat band pattern in gel electrophoresis reminiscent of the nuclease digestion pattern of chromatin, with monosome-size fragments as the digestion limit. The H2cyt141-induced nucleolytic effect would therefore occur in the linker regions of cell DNA and might result from the observed stimulation (by a factor of greater than 100) of an acidic (optimum pH 4.0) endonuclease activity. The nucleolytic effect also appeared to be recessive in vitro and absent in mixed samples containing extracts from H2cyt141-infected cells plus extracts from H2WT- or mock-infected cells. The virus gene product responsible for the enhancement of the acidic endonuclease was found to function stoichiometrically and not catalytically. The cytocidal and nucleolytic effects of the viral E1B region 19K protein may be mediated by a cellular inhibitor of acidic endonuclease.

Adenoviruses, Human↗
PubMed1984

Initiation rate of adenovirus DNA synthesis in infected cell.

A method was developed to determine the rate of viral DNA synthesis initiation in adenovirus 2-infected cells. The initiation of DNA synthesis appeared as the rate-limiting step for accumulation of viral DNA. The multiplicity of infection slightly influenced the rate of synthesis of viral DNA, and only during the linear phase of viral DNA production. The initiation of DNA-synthesis was found to occur preferentially on newly synthesized DNA molecules. These kinetics data and the effect of novobiocin suggested that binding of viral DNA with some enzymatic complexes favored the replication of a minor, active class of adenovirus DNA molecules.

Adenoviruses, Human↗
PubMed1984

Genetic expression of human adenoviruses in simian cells. Evidence for interserotypic inhibition of viral DNA synthesis.

Most simian cells are permissive for SV40 and adenovirus-SV40 hybrids but nonpermissive for human adenoviruses, and the defect has been shown to take place at the level of processing of late viral mRNAs (Klessig and Grodzicker, 1979). Viral DNA synthesis and virus progeny production were studied in simian cells infected with different adenovirus serotypes. Adenoviruses belonging to oncogenic subgroups A and B (Ad31 and Ad3) failed to replicate their DNA in CV1 cells, whereas DNA replication occurred for all the other serotypes. Co-infection of CV1 cells with SV40 and Ad3 (or Ad31) resulted in the inhibition of SV40 DNA synthesis, as well as cellular DNA synthesis. The inhibition was not related to adenovirus DNA replication, since SV40 did not complement the Ad3/Ad31 replication defective function. Similar results were obtained in coinfected BSC and MK2 simian cell lines. Inhibition of Ad2ND1 DNA synthesis and gene expression also occurred in co-infection of simian cells with nondefective Ad2ND1 hybrid and defective Ad3/Ad31. In permissive human cell lines (HeLa or KB) co-infected with Ad2 and Ad3 (or Ad31), a dominant, inhibitory effect of Ad3 (or Ad31) over Ad2 was also observed. The inhibition appeared to function stoichiometrically and not catalytically, and to involve early adenovirus gene products. In both simian and human cells a hierarchy of dominance appeared between serotypes belonging to different subgroups. The degree of inhibitory effect occurred in the following decreasing order: Ad3 and Ad7 (subgroup B), Ad9 (D), Ad4 (E), Ad31 (A), Ad2 and Ad5 (C).

Adenoviruses, Human↗
PubMed1984

Restriction maps of human adenovirus types 2, 5, and 3 for BstEII, MluI, NdeI, NruI and SfiI endonucleases.

The positions of cleavage sites for BstEII, MluI, NdeI, NruI and SfiI restriction endonucleases in the DNA from human adenovirus (Ad) serotypes 2, 5 and 3 were determined. In addition, the sites of cleavage for BglII in Ad3 DNA were located. All these enzymes possess a narrow specificity and generated a small number of discrete DNA fragments. Ad3 DNA was not cleaved by MluI and SfiI. It was the first observation of the absence of cleavage of an adenovirus DNA by a restriction endonuclease.

Adenoviruses, Human↗
PubMed1984

Characterization of an early temperature-sensitive and cytocidal double mutant of adenovirus type 2.

Human adenovirus type 2 mutant, H2 ts 111, presented a double phenotype: temperature-sensitive (ts) for initiation and elongation of DNA synthesis, and cytocidal (cyt) by its large-plaque formation and the nucleolytic cleavage of both viral and cellular DNAs. Both characters were recessive since they were efficiently complemented by wild-type or other DNA-negative ts mutants. H2 ts 111 DNA--terminal protein complex formed at 33 degrees C and chased at 39.5 degrees C showed a decreased affinity for glass fibre filters, concurrently with the loss of protein-linked DNA ends. H2 ts 111 DNA breakdown occurring upon shift-up to 39.5 degrees C therefore appeared to start in close proximity to the genome extremities. Marker rescue experiments showed that the ts character was abolished by co-infection with plasmid recombinants containing whole or part of the E2A region, encoding for the 72K DNA-binding protein. The N-terminal domain of this 72K protein has been assigned between 0 and 200 amino acids, and is supposed to have a function in late transcription control. Since H2 ts 111 mapped between 0 and 300 residues (63.6 to 65.9 map units), its mutation was most likely located between 200 and 300 amino acids, namely in the C-terminal domain of the protein, which is involved in DNA replication. Recombination between H2 ts 111 and H5 dl 313 mutant revealed that the cyt function was localized in the E1B zone, between 3.8 and 11.3 map units. The nucleolytic and cytocidal effects were complemented by HEK 293 cells, an H5-transformed cell line expressing the left-most 12.5% of the viral genome. H2 ts 111 appeared, therefore, as a double ts-cyt mutant. The gene product rendered temperature-sensitive by the H2 ts 111 mutation was found to act stoichiometrically, and not catalytically, a result compatible with a lesion in the 72K protein. Although inactive at the non-permissive temperature, the early 72K protein was normally synthesized and stable in H2 ts 111-infected cells at 39.5 degrees C. Assymetric complementation obtained with the DNA-defective H5 ts 36 implied a certain degree of type specificity in the DNA-binding protein function.

Adenoviruses, Human↗
PubMed1982

Physical mapping of adenovirus type 2 temperature-sensitive mutations by restriction endonuclease analysis of interserotypic recombinants.

The genome structures of about 100 interserotypic ts recombinants produced in crosses between human adenovirus type 2 (H2) and 5 (H5) temperature-sensitive mutants were analyzed by cleavage with restriction endonucleases to determine the map coordinates of the following temperature-sensitive mutants: penton base plus fiber-defective H2 ts103, -104, and -136, assembly-defective H2 ts112, fiber-defective H2 ts125, hexon-defective H2 ts118 and -121, and DNA-negative H2 ts111. H5 ts1 (100 K defective), H5 ts36 (DNA negative), H5 ts125 (mutated in the early 72,000-dalton protein), H5 ts22 (fiber defective), H5 ts58 (IIIa defective), and H5 ts18 and -19 were used as one of the parents. The physical locations of the H2 temperature-sensitive mutations thus defined are discussed in relation to the genetic map, the biological function altered, and the positions of the structural genes on the genome.

Adenoviruses, Human↗
PubMed1980

Effects of novobiocin on adenovirus DNA synthesis and encapsidation.

Novobiocin, an inhibitor of DNA gyrase implicated in bacterial and likely mammalian, chromosome replication, inhibited the initiation, but not the elongation of human adenovirus DNA replicative synthesis. The inhibition was partially reversible, even in the presence of protein synthesis inhibitor. Novobiocin inhibited also the encapsidation of viral DNA, and this effect was independent of the block in DNA replication. It was suggested that novobiocin acted on two different functions, one involved in viral DNA replication initiation, the other in DNA encapsidation.

Adenoviruses, Human↗
PubMed1980

Morphogenesis of human adenovirus type 2 studied with fiber- and fiber and penton base-defective temperature-sensitive mutants.

The nature, polypeptide composition, and antigenic composition of the particles formed by six human adenovirus type 2 temperature-sensitive (ts) mutants were studied. ts115, ts116, and ts125 were phenotypically fiber-defective mutants, and ts103, ts104, and ts136 failed to synthesize detectable amounts of fiber plus penton base at 39.5 degrees C. The mutants belonged to five complementation groups, one group including ts116 and ts125. Except for ts103 and ts136, the other mutants were capable of producing particles at 39.5 degrees C. ts116 and ts125 accumulated light assembly intermediate particles (or top components) at nonpermissive temperatures, with few virus particles. The sodium dodecyl sulfate polypeptide pattern of ts116- or ts125-infected cells, intermediate particles, and virus particles showed that polypeptide IV (fiber) was smaller by a molecular weight of 2,000 than that in the wild-type virion and was glycosylated. In fiber plus penton base-defective ts104-infected cells, equivalent quantities of top components and viruses with a buoyant density (rho) of 1.345 g/ml (rho = 1.345 particles) were produced at 39.5 degrees C. These rho = 1.345 particles corresponded to young virions, as evidenced by the presence of uncleaved precursors to proteins VI, VIII, and VII. These young virions matured upon a shift down. Virus capsid vertex antigenic components underwent a phase of eclipse during their incorporation into mature virus particles. No antigenic penton base or IIa was detected in intermediate particles of all the ts mutants tested. Only hexon and traces of fiber antigens were found in ts104 young virions. Penton base and IIIa appeared as fully antigenically expressed capsid subunits in mature wild-type virions or ts104 virions after a shift down. The ts104 lesion is postulated to affect a regulatory function related in some way to penton base and fiber overproduction and the maturation processing of precursors PVI, PVII, and PVII.

Adenoviruses, Human↗
PubMed1979

Adenovirus early function required for protection of viral and cellular DNA.

Studies were done to characterize a DNA-negative temperature-sensitive (ts) mutant of human adenovirus type 2, H2 ts111. The temperature-sensitive defect, which was reversible on shift-down in the absence of protein synthesis, was expressed as early as 2 h postinfection, and the results of density-labeling experiments are in agreement with at least a DNA replication initiation block. On shift-up, after allowing viral DNA synthesis at permissive temperatures, the newly synthesized viral DNA and the mature viral DNA were cleaved into fragments which sedimented as a broad peak with a mean coefficient of 10-12S. This cleavage was more marked in the presence of hydroxyurea as the DNA synthesis inhibitor. Parental DNA in infected cells was degraded to a much lesser extent regardless of the incubation temperature. In contrast, the parental DNA was strongly degraded when early gene expression was permitted at 33 degrees C before shift-up to 39.5 degrees C. Furthermore, cellular DNA was also degraded at 39.5 degrees C in ts111-infected cells, the rate of cleavage being related to the multiplicity of infection. This cleavage effect, which did not seem to be related to penton base-associated endonuclease activity, was also enhanced when early gene expression was allowed at 33 degrees C before shift-up. The ts111 defect, which was related to an initiation block and endonucleolytic cleavage of viral and cellular DNA, seemed to correspond to a single mutation. The implication of the ts111 gene product in protection of viral and cellular DNA by way of a DNase-inhibitory function is discussed.

Adenoviruses, Human↗
PubMed1978

Isolation and phenotypic characterization of human adenovirus type 2 temperature-sensitive mutants.

Thirty-nine temperature-sensitive (ts) mutants that fail to grow at 39.5 degrees C but develop normally at 33 degrees C have been isolated from a nitrous-acid-treated stock of a wild-type strain of type 2 human adenovirus. The frequency of ts mutants among the surviving viruses was about 10%. Complementation tests in doubly infected cell cultures at restrictive temperature permitted the assignment of 19 of these mutants to 11 complementation groups. They are characterized phenotypically according to their soluble capsid antigen production quantified by two-dimensional immunoelectrophoresis, virus DNA synthesis, as measured by alkaline sucrose gradient sedimentation of 34S DNA, and virion morphogenesis, as analysed by electron microscopy os cell sections. Two complementation groups were defective for DNA synthesis, four for soluble hexon production and two groups for total penton (penton base + fibre), while one group revealed no fibre production. Two complementation groups presented a normal antigen pattern, but the particles exhibited altered morphology as observed in cell sections.

Adenoviruses, Human↗