PubMed Health⌕ Search

Biomedical subjects

A Gazit

Publications and source records attributed to A Gazit.

At least 127 records · Page 7Linked to original sources

Nucleotide sequence analysis of equine infectious anemia virus proviral DNA.

The nucleotide sequence of the integrated form of the genome of the equine infectious anemia virus was determined. By comparison with LTR sequences of other retroviruses, signals for the control of viral gene transcription and translation could be identified in the EIAV LTR. Open reading frames for gag and pol genes were identified and their sequences matched very closely to those determined previously by others. However, in the present study, the pol gene reading frame was open throughout its entire length. The open reading frame for the env gene product was constructed from the sequences of two independent EIAV clones. Thus, a noninfectious genomic-length clone was shown to contain a frameshift mutation approximately in the middle of the presumed env gene coding sequence, whereas the sequence of another clone was open in this region. The deduced amino acid sequences of the EIAV gag and pol products showed closer evolutionary relationships to those of known lentiviruses than to other retroviruses. There was also partial sequence homology between predicted env gene products of EIAV, visna virus, and HTLV-III/LAV. Sequences analogous to the sor region of other lentiviruses could not be identified in our EIAV clone. A short open reading frame at the 3' end of the genome that overlapped env but not the 3' LTR was present but lacked significant sequence similarity to the 3' open reading frames of other lentiviruses. Thus, the sequence and general structure of EIAV most closely resemble those of known lentiviruses.

Amino Acid Sequence↗

Molecular cloning and physical characterization of integrated equine infectious anemia virus: molecular and immunologic evidence of its close relationship to ovine and caprine lentiviruses.

Molecular clones of the integrated form of the genome of equine infectious anemia virus (EIAV), the etiologic agent of a naturally occurring, worldwide disease of horses, were obtained. The restriction map of a full-length genome was determined. Additional evidence for the close evolutionary relationship between EIAV and a prototype lentivirus (caprine arthritis encephalitis virus) was acquired by Southern blotting and immunological analyses. An interspecies radioimmunoassay was developed in which EIAV and ovine and caprine lentiviruses could be detected equally well. These studies make available precisely defined reagents to pursue the study of the mechanisms of pathogenesis of lentiviral induced diseases.

Animals↗

Analysis of structural polypeptides of the lymphoproliferative disease virus (LPDV) of turkeys.

The polypeptide composition of the lymphoproliferative disease virus (LPDV) of turkeys was shown to comprise several polypeptides with apparent molecular weights of 76, 31, 28, 20 and 15 kDa. This polypeptide pattern is distinctly different from the protein profiles of avian leukosis viruses, reticuloendotheliosis virus, or murine leukemia viruses. Moreover, LPD virions contain 2 major structural proteins (p31 and p28), in contrast to only one major internal protein present in most other retroviruses. The 76 kDa protein was established as the major viral envelope glycoprotein. The uniqueness of the LPDV polypeptide pattern is consistent with the lack of genetic relatedness of LPDV genome to other retroviruses, establishing LPDV as a representative of a distinct group of retroviridae.

Animals↗

Nucleotide sequence analysis of the long terminal repeat of integrated caprine arthritis encephalitis virus.

The nucleotide sequence of the long terminal repeat (LTR) of caprine arthritis encephalitis virus (CAEV), a prototype lentivirus was determined. 6-bp directly repeated host cell sequences flank the 376-bp proviral LTRs. By comparison with other retroviral sequences, the CAEV LTR likely contains U3, R and U5 regions 207, 86 and 83 base-pairs in length, respectively. Sequences conforming to consensus transcriptional promoter sites were identified in the U3 region upstream of a potential transcription initiation site. A consensus polyadenylation signal is present 20 bases upstream of the putative R-U5 border and a potential poly(A) addition site. Sequence comparisons of the CAEV LTR with those of other retroviruses uncovered significant similarities with that of visna virus. No other global homologies with other retrovirus LTRs could be detected. CAEV utilizes a primer binding site complementary to lysine tRNA as does visna, AIDS associated retroviruses, and mouse mammary tumor virus. The putative primer for positive-strand DNA synthesis identified in the CAEV sequence is identical to that of visna virus and very similar to those of AIDS retroviruses and MMTV. In addition, a stretch that includes the TATA box of the CAEV LTR resembles closely the corresponding region in the AIDS retrovirus. These and other findings further strengthen the classification of AIDS retrovirus as a lentivirus.

Animals↗

Mammalian cell transformation by a murine retrovirus vector containing the avian erythroblastosis virus erbB gene.

A recombinant murine retrovirus vector containing the v-erbB gene of avian erythroblastosis virus was constructed to investigate v-erbB as a transforming gene for mammalian cells. A restriction fragment containing the v-erbB sequences from a molecular clone of avian erythroblastosis virus was inserted into a Moloney murine leukemia virus vector. The construct, designated MuLV/erbB, transformed NIH 3T3 cells at a high efficiency in the DNA transfection assay. Individual MuLV/erbB transfectants grew in soft agar and were tumorigenic. The transfectants contained v-erbB DNA sequences, expressed v-erbB-specific transcripts, and synthesized v-erbB-related glycoproteins. The majority of transfectants produced two major v-erbB gene products of 58 and 66 kilodaltons. However, some transfectants produced much smaller v-erbB-specific proteins. Tunicamycin experiments revealed that the size heterogeneity observed between different transfectants was not due to variations in glycoprotein processing, implying that, in some cases, alterations in the MuLV/erbB genome occurred during the transfection process. These findings indicate that expression of the complete v-erbB gene product is not required for transformation of NIH 3T3 cells. A transmissible murine v-erbB (M-erbB) virus was generated by infection of nonproducer transfectants with amphotrophic murine leukemia virus. Transmission of the rescued M-erbB virus was confirmed by DNA, RNA, and protein analyses. The introduction of a transforming v-erbB gene into mammalian cells by virus infection provides a means of analyzing the mechanism by which this epidermal growth factor receptor-related gene alters the growth and differentiation of cells from various lineages.

Alpharetrovirus↗

Synthesis and receptor-binding affinity of fluorotamoxifen, a possible estrogen-receptor imaging agent.

Aminotamoxifen was totally synthesized from p-nitrobenzoyl chloride via a Friedel-Crafts acylation. Then, by means of a Balz-Schiemann reaction, aminotamoxifen was converted into fluorotamoxifen. The triazene variation of this conversion, with a 25% yield, enables a rapid, one-step diazotization, incorporating a fluorine atom into the phenyl ring of the tamoxifen. This reaction may be useful for the preparation of low specific activity 18F-labeled tamoxifen, for distribution, and for estrogen-receptor studies. For these in vivo and in vitro studies, fluorotamoxifen was also synthesized from p-fluorobenzoyl chloride, and its chemical intermediates were compared with estradiol and hexestrol, for their receptor binding and competition, as well as for their uterotropic activity. It is demonstrated that tamoxifen and fluorotamoxifen are strong estradiol agonists and partial hexestrol agonists, while aminotamoxifen is a weak estradiol and hexestrol agonist.

Animals↗

Iron, transferrin, and acid and alkaline phosphatase in healthy turkeys and in turkeys inoculated with the lymphoproliferative disease virus.

Presented are data on iron-binding capacity determinations in the serum of turkeys infected with lymphoproliferative disease (LPD) virus and in healthy males and females (laying eggs and nonlaying) from a breeding flock. Also presented are results of serum and tissue total acid and alkaline phosphatase determinations in turkey poults infected with LPD virus and their uninfected controls and of serum enzyme levels in healthy males and females from the breeding flock. There was no significant alteration in total iron binding capacity (transferrin level) in the serum of turkeys with LPD. Turkey poults inoculated with LPD virus showed a significant decrease in serum alkaline phosphatase activity 4 and 7 weeks postinfection (pi), and a decrease in serum acid phosphatase activity 7 weeks pi. Acid and alkaline phosphatase activity determined in the spleen and pancreas (organs with pronounced tumor involvement) 7 weeks pi did not differ significantly from that of healthy controls, although there was a tendency for both enzymes to decline in the pancreas of the infected turkeys. Healthy laying female turkeys demonstrated marked elevation in serum transferrin level and in acid and alkaline phosphatase activity, as compared with males of the same age. Serum alkaline phosphatase of turkey poults was markedly higher than that of adult turkeys.

Acid Phosphatase↗

Appearance of ovine progressive pneumonia virus outside the United States.

A recently isolated Israeli retrovirus from a sheep with maedi-visna was compared with other retroviruses, using cDNA-RNA hybridization in solution. The Israeli isolate was shown to have close, if not identical, genetic homologic features with the ovine progressive pneumonia virus reported in the United States, rather than with those of the maedivisna viruses of European origin.

Animals↗

Isolation and characterization of a novel retrovirus from sheep affected by pulmonary carcinoma.

Sheep pulmonary carcinoma (SPC) has been shown to be associated in nature with a retrovirus, by electron microscopic, biochemical, and epidemiological criteria and by experimental transmission. In this study, a retrovirus has been isolated from SPC tumors which were experimentally induced by inoculation with a cell-free, reverse transcriptase containing fraction from a spontaneous field case of SPC, and propagated in culture. This novel virus was shown to be unrelated to murine, avian, and bovine leukemia viruses, to be exogenous to the ovine species, and to have only limited genetic relatedness to the lentiviridae (maedi-visna and caprine arthritis encephalitis virus).

Animals↗

Expression of the normal human sis/PDGF-2 coding sequence induces cellular transformation.

The human sis proto-oncogene contains the coding sequence for one of two polypeptide chains present in preparations of biologically active human platelet-derived growth factor (PDGF). A human clone, c-sis clone 8, which contains all of the v-sis-related sequences present in human DNA, was transcriptionally inactive when transfected into NIH/3T3 cells. When placed under the control of a retrovirus LTR, the clone was transcribed at levels comparable to that observed in cells transformed by SSV DNA. However, c-sis clone 8 DNA did not express detectable sis/PDGF-2 proteins and lacked biologic activity. A putative upstream exon was identified by its ability to detect the 4.2 kb sis-related transcript in certain human cells. When this sequence was inserted in the proper orientation between the LTR and c-sis clone 8, the chimeric molecule acquired high titered transforming activity, comparable to that of SSV DNA. Transformants containing this construct expressed human sis/PDGF-2 translational products. Thus the normal coding sequence for a human growth factor has transforming activity when expressed in an appropriate assay cell.

Amino Acid Sequence↗

Genetic control of the organ specificity of lymphoproliferative disease virus (LPDV) of turkeys.

In a previous study based on the kinetics of virus replication and tumor formation (Gazit et al., 1982), it was shown that the organotropism of lymphoproliferative disease virus (LPDV) is confined to lymphoid tissues. The present paper demonstrates that this organ specificity is controlled at the level of infection and integration, that is, the lymphoid organs, which are the only organs sustaining virus replication, and also the only organs in whose cells integrated LPDV proviruses are detectable. At the same time, the efficiency of virus replication within the various target organs is regulated both at the level of infection and integration and at the level of viral gene transcription.

Animals↗

The caprine arthritis-encephalitis virus is a distinct virus within the Lentivirus group.

The genetic relatedness among viral genomes of caprine arthritis-encephalitis virus, visna virus, and progressive pneumonia virus, was determined. Whereas the genomic RNAs of two strains of visna virus are indistinguishable as reflected both by their annealing kinetics as well as by the thermal stability of the hybrids, the caprine arthritis-encephalitis virus and visna virus have only 30% of their nucleic acid sequences in common. Furthermore, within the homologous regions of the two viral genomes, there is a significant level (approximately 10%) of mismatched base pairs. This limited homology that exists between caprine arthritis-encephalitis virus and visna virus was lower than the sequence homology observed between the genomes of visna virus and progressive pneumonia virus, or between the genomes of caprine arthritis-encephalitis virus and progressive pneumonia virus. All this indicates that caprine arthritis-encephalitis virus is an additional distinct member of the Lentivirus group of the Retroviridae family.

Animals↗

Organotropism of the lymphoproliferative disease virus (LPDV) of turkeys.

Turkey poults were inoculated with lymphoproliferative disease virus (LPDV) of turkey, and the organotropism of the inoculated virus was determined from the kinetics of virus expression and tumor formation. Molecular hybridization experiments, conducted to determine the level of viral RNA expression in the various organs of infected birds, established lymphoid tissues, including bone marrow, as the target for LPDV infection. Of these, bone marrow was the first to be infected but subsequently virus replication extended to the thymus, the spleen and the bursa Of Fabricius. The low level of LPDV expression in non-target organs probably stemmed from infiltration of infected lymphatic cells. The fact that the main organs for virus replication were not invariably the ones most clinically involved suggests that the target cells for virus infection need not necessarily be the targets for virus oncogenicity. A possible mode for LP[DV infection and transformation is proposed.

Animals↗

DNA-processing activities associated with the purified alpha, beta 2, and alpha beta molecular forms of avian sarcoma virus RNA-dependent DNA polymerase.

The RNA-dependent DNA polymerase purified from B77 avian sarcoma virus exhibited two distinct DNA-processing activities. The alpha and beta 2 isoenzymes possessed an endodeoxyribonuclease activity capable of nicking simian virus 40 superhelical DNA, whereas the alpha beta isoenzyme performed as an untwisting topoisomerase. Both activities associated with the three molecular forms of the retroviral DNA polymerase were dependent on the presence of either Mn2+ or Mg2+ ions. From analysis of the denaturated DNA products, it is apparent that the alpha and beta 2 isoenzymes introduced two nicks, one per each strand in the superhelical simian virus 40 DNA molecules, whereas the alpha beta polymerase converted these supercoiled molecules to the relaxed covalently closed circular form. The notion that the DNA-processing activities are located on the DNA polymerase molecules was supported by the following: (i) the three isoenzymes were of a high purity; (ii) the activities cosedimented in glycerol gradients with the DNA polymerase activities of the alpha, beta 2, and alpha beta molecular forms; and (iii) immunoglobulin directed against the purified polymerase immunoprecipitated the DNA-processing activities. Chemical treatments of the DNA polymerase molecules (with pyridoxalphosphate, iodoacetamide, and sulfhydryl reagents), which inhibited the polymerase activity, also suppressed the endonucleolytic and topoisomerase activities, suggesting that cystein and amino groups play an important role in the active sites of the DNA-processing activities as well.

Alpharetrovirus↗

Host range and oncogenicity studies of the murine myeloma MOPC-315 type-C retrovirus.

The biological features of an endogenous type-C RNA tumor virus released by the murine plasmacytoma MOPC-315 were determined. The virus was found to be pure ecotropic and, according to its capability to infect both NIH/3T3 and BALB/3T3 cells, it was established as an NB tropic virus. This tropism became more pronounced as a consequence of multiple passage in balb/3T3 cells. In view of the ability of myeloma viruses to efficiently infect BALB/c cells, their possible role in myeloma induction is considered.

Animals↗