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Y Cleuter

Publications and source records attributed to Y Cleuter.

At least 37 records · Page 2Linked to original sources

Role of the 3' long open reading frame region of bovine leukemia virus in the maintenance of cell transformation.

Viral RNA expression was studied by dot blot hybridization with polyadenylated RNAs extracted from a bovine (YR-1) and an ovine (YR-2) tumor cell clone. Both clones were derived from in vivo bovine leukemia virus-induced tumors. The probes used were either the bovine leukemia virus information or only the long open reading frame sequences. No viral RNA corresponding to the bovine leukemia virus long open reading frame region was detected in YR-2, and a very limited amount of bovine leukemia virus messages was unraveled in YR-1. These results strongly suggest that viral expression, even in the long open reading frame region, is not required to maintain transformation of at least some tumor cells.

Animals↗

Bovine leukemia virus, a versatile agent with various pathogenic effects in various animal species.

The bovine leukemia virus is the etiological agent of a chronic lymphatic leukemia in cows, sheep, and goats. The same virus seems to induce a kind of wasting disease in experimentally infected rabbits. Antibodies to highly purified bovine leukemia viral Mr 51,000 glycoprotein and Mr 24,000 protein cross-react with human T-lymphotropic virus III/lymphadenopathy-associated virus antigens present in cultured lymphocytes of African patients suffering from acquired immune deficiency syndrome. Bovine leukemia virus has many structural and functional characteristics in common with the human T-lymphotropic viruses. The most striking feature of these retroviruses is the existence of a long open reading frame located at the 3' side of the provirus between the right end of the 3' side of env gene and the left end of the long terminal repeat. It is believed that the long open reading frame protein product acts in trans upon a number of genes to account for the biological effects of the virus.

Animals↗

Bovine leukaemia virus and enzootic bovine leukosis.

Infection of bovines with bovine leukaemia virus (BLV) manifests itself in either of two ways: 30-70% of carriers develop persistent lymphocytosis (PL), with the viral genome integrated at a large number of different sites in the DNA of the affected B-lymphocytes, without causing any chromosomal abnormalities. Only 0,1-10% of carriers develop lymphoid tumours, which also consist of B-lymphocytes. In contrast to PL, however, they are of mono- or oligoclonal origin in terms of the integration site, which is characteristic for each tumour. All cells contain one or more copies of the viral genome, chromosomal aberrations are common and if deletions are present they are invariably found in the 5'-half of the virus DNA sequence. In both types of affected cells transcription is repressed in vivo, but transient virus production can be induced in vitro and detected by means of syncytia induction or haemagglutination. In vivo production of virus in some unknown cell is suggested by the presence of high antibody titres in infected animals, especially against the envelope glycoprotein gp51. This can be detected by various techniques such as immunodiffusion, radioimmune assay or ELISA. Monoclonal antibodies against gp51 have revealed 8 epitopes, 3 of which are recognized by neutralizing antibodies and one by a cytolytic antibody. The BLV genome, about 9 kb in size, have been cloned, and some of the information obtained on its molecular structure and function is discussed. It codes for at least 4 non-glycosylated and 2 glycoproteins. Of special interest is the recently discovered serological relationship between some of the non-glycosylated proteins and those of the human T-cell leukaemia virus. The functional role of BLV in leukaemogenesis is largely unknown. The presence of the viral genome seems to be necessary for the maintenance of the transformed state, but not its continuous expression nor an LTR-mediated promotion of transcription of cellular genes. No oncogene is carried by the virus. Although bovine leukosis is not of major economic importance, its eradication is desirable and feasible in countries with a relatively low incidence, by means of testing and elimination. For endemic situations vaccination would be preferable, and distinct possibilities exist for the development of gp51 based vaccines.

Animals↗

Biologically active epitopes of bovine leukemia virus glycoprotein gp51: their dependence on protein glycosylation and genetic variability.

A panel of monoclonal antibodies to the bovine leukemia virus envelope glycoprotein (BLV gp51) has previously demonstrated the association of the biological activities of the virus (infectivity, syncytia induction) with three out of eight epitopes of gp51. In BLV-infected cells, the unglycosylated homolog of the precursor to the BLV envelope glycoproteins (gPr72env) is a 47,000-MW polypeptide. Immunoprecipitation studies with monoclonal antibodies show that the neutralizing antibody-inducing sites, although present in gPr72env, are not conserved in the 47,000-MW unglycosylated homolog. Finally, it is demonstrated that the neutralizing antibody-inducing sites of gp51 are subject to antigenic variation among BLV isolates of the same or different geographical origins.

Animals↗

Ultrastructural demonstration of retrovirus antigens with immuno-gold staining in prodromal acquired immune deficiency syndrome.

The peripheral lymphocytes of a patient with prodromal acquired immune deficiency syndrome contained giant multivesicular bodies. These were specifically stained by immuno-gold labelled polyclonal antibodies against the major core protein p24 of bovine leukaemia virus and human T cell leukaemia virus I. Moreover, the patient's serum was positive for bovine leukaemia virus by the ELISA method.

Acquired Immunodeficiency Syndrome↗

Bovine leukemia virus, a distinguished member of the human T-lymphotropic virus family.

Bovine leukemia virus (BLV) has many structural and functional characteristics in common with the human T-lymphotropic viruses (HTLVs). The most striking feature of these retroviruses is the existence of a long open reading frame (LOR) located at the 3' side of the provirus between the right end of the 3' side of env gene and the left end of the long terminal repeat (LTR). It is believed that the LOR protein product is of critical importance in the induction process of the tumor phase of bovine leukemia. Prevention of BLV infection will be attempted by vaccination. To that aim, careful study of BLV envelope glycoprotein epitopes has shown that epitopes F, G, and H play a major role in biological activities of the virus. Their native structure depends upon glycosylation of the peptide backbone.

Animals↗

[Oncogenes].

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Animals↗

Translational stability of plant viral RNAs microinjected into living cells. Influence of a 3'-poly(A) segment.

Three different alternative structural features have been shown to be present at the 3' terminus of plant viral RNAs: (a) a poly(A) track, (b) a tRNA-like structure, (c) no special structural or sequence characteristic. We have compared the translational stability after injection into frog oocytes of a representative of each type: (a) the small genomic RNA (M-RNA) of cowpea mosaic virus (CPMV), (b) the subgenomic mRNA for coat protein (RNA 4) of brome mosaic virus (BMV), (c) the subgenomic mRNA for coat protein (RNA 4) of alfalfa mosaic virus (AIMV). It has been shown that CPMV M-RNA exhibits the highest translational stability. However, the stability of AIMV RNA 4 is remarkably high and moreover significantly higher than that of BMV RNA 4. We demonstrate that, for all three viral RNA species considered, the presence of a poly(A) segment at the 3' end of the molecules improves the translational stability. From a comparative investigation in which AIMV RNA 4 was also injected into HeLa cells, it is concluded that the stability of a given non-adenylylated mRNA depends on the nature of the cytoplastic environment.

Animals↗

Leukemogenesis by bovine leukemia virus: proviral DNA integration and lack of RNA expression of viral long terminal repeat and 3' proximate cellular sequences.

The DNA from 17 lymphoid tumors induced by bovine leukemia virus (BLV) was digested with the restriction endonuclease EcoRI. Filter hybridization analysis using radioactive probes specific for the BLV genome showed that all tumors contained at least one or a portion of one provirus. Digestion of these proviruses with Sac I demonstrated that deletions occurred in about 25% of the cases and involved sequences located in the 5' half of the provirus. No sequence homology was observed between the cloned proximate cellular sequences flanking two different proviruses at their 3' end and the corresponding sequences in 16 other tumor DNAs, thus showing that a wide range of genomic sites could accommodate BLV proviruses. Transcription of viral DNA including long terminal repeated sequences was not detected, strongly suggesting that viral gene expression is not required for maintenance of the tumor state. No expression of 3'-proximate cellular sequences was observed, indicating that no proximate downstream promotion took place in the cases examined.

Animals↗

Viral RNA content of bovine leukemia virus-infected cells.

A bovine leukemia virus (BLV)-producing cell line, fetal lamb kidney cells infected with BLV (FLK) contains one or a few copies of BLV proviral DNA in its genome. These cells contain 0.002% of viral RNA which sediments, in a sucrose gradient, at about 35S and between 18S and 28S. In cattle affected by enzootic bovine leukosis, tumor cells and circulating lymphocytes also contain one or a few copies of BLV proviral DNA integrated in their genome. However, in all cases tested (except one), no viral RNA was detected in these cells in conditions where one or two copies of viral genomic RNA per cell would have been easily detected.

Animals↗

Effect of tRNA pool balance on rate and uniformity of elongation during translation of fibroin mRNA in a reticulocyte cell-free system.

Unsuccessful attempts to synthesize complete fibroin chains in vitro were previously made in heterologous cell-free system [3]. In the present work, we succeeded to obtain complete translation of purified fibroin mRNA in a rabbit reticulocyte lysate. Whilst this work was being completed [1], similar results were published by Lizardi et al. [4]. The synthesis of full-sized molecules of fibroin (M.W. 360,000) was achieved by adding tRNA from the posterior silk gland to the cell-free system. With tRNA from other sources, both the translation rate and the amount of complete fibroin chains dropped. This effect of tRNA is situated at the elongation levels. Analysis of cell-free synthesized products by polyacrylamide gel electrophoresis shows that smaller discrete polypeptides are accumulated after 120 minutes of incubation. These polypeptides correspond to growing fibroin chains. This pattern of translation products suggests that elongation might decelerate at specific sites of the fibroin mRNA. These results show that a tRNA pool adjusted to mRNA codon frequency is required to obtain the maximal average elongation rate. A stochastic model based on random acceptance of tRNA at the ribosomal A site for the codon-anticodon recognition process can explain this phenomenon. It can also explain the occurrence of the unfinished discrete fibroin polypeptides during in vitro translation.

Animals↗

Translational stability of native and deadenylylated rabbit globin mRNA injected into HeLa cells.

HeLa human cells were injected with a natural mixture of rabbit alpha and beta globin mRNA. They were incubated for 6 hr with [35S]methionine either immediately after injection or 20 hr later. The labeled proteins in the injected cells were analyzed by fluorography of two-dimensional electrophoresis gels. By using this procedure, it was possible to show that, during the first few hours after injection, both alpha and beta globin molecules are synthesized with an alpha to beta ratio approximately equal to 0.6. The rate of synthesis of alpha globin decreased significantly faster than that of beta globin over a 26-hr period after injection of the two mRNAs. It thus seems that two messenger RNAs coding for closely related polypeptides possess a markedly different translational stability. When deadenylylated rabbit globin mRNAs were injected into HeLa cells, no globin synthesis could be detected by the techniques used. We conclude that the translational half-life of mRNAs lacking poly(A) is very short in these cells. It is thus clear that the poly(A) segment is required to ensure stability to globin mRNA in somatic cells as in Xenopus oocytes.

Adenine↗

Genomic integration of bovine leukemia provirus: comparison of persistent lymphocytosis with lymph node tumor form of enzootic.

Integration of bovine leukemia proviral DNA in the genome of infected cells was investigated in cattle affected by either the persistent lymphocytosis or the lymph node tumor form of enzootic bovine leukosis. In persistent lymphocytosis, proviral DNA was found to be integrated at a large number of genomic sites in one-fourth to one-third of circulating leukocytes. In the lymph node tumor form, in contrast, proviral DNA was found to be integrated at one or very few sites in the genomes of a larger fraction of both circulating leukocytes and lymph node tumor cells.

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Humoral antibody response to bovine leukemia virus infection in cattle and sheep.

In this study, 345 cattle from 7 herds with a history of lymphosarcoma were tested for antibody to BLV antigens by three serological methods, namely immunodiffusion using a bovine leukemia virus glycoprotein with a molecular weight of 60,000 as antigen, and radioimmunoassay using a bovine leukemia virus glycoprotein with a molecular weight of 60,000 and a bovine leukemia virus protein with a molecular weight of 24,000 as antigen. The three tests under comparison agreed for 335 animals, 240 being negative in the three tests, and 95 being positive. Results were variable in ten cases only. Glycoprotein with a molecular weight of 60,000 antibody titers were systematically higher than were protein with a molecular weight of 24,000 antibody titers in bovine sera and milk, as well as in sera of experimentally infected sheep. In the latter case, antibodies to bovine leukemia virus antigens reached maximal values at the animal death in the tumor phase of the disease. Ratios of serum antiglycoprotein titer to milk titer varied between 4 and 117, showing that, if milk pools are to be used in surveys of bovine leukemia virus infection, use of very sensitive techniques of detection is mandatory.

Animals↗