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Y Shaul

Publications and source records attributed to Y Shaul.

66 records · Page 4Linked to original sources
PubMed1985

A human hepatitis B viral enhancer element.

Fragments of the cloned hepatitis B virus (HBV) genome were assayed in vivo for the presence of a transcriptional enhancer element. We demonstrate that sequences positioned approximately 450 bp upstream from the HBcAg gene promoter are required for its efficient activity. These HBV stimulatory sequences activate transcription when inserted upstream to a heterologous SV40 early promoter. Like other known enhancer elements, this HBV sequence acts in an orientation-independent manner. Furthermore, the HBV enhancer element exhibits a preferred activity in a human hepatoma cell line.

Animals↗
PubMed1985

Homologous recombination between a defective virus and a chromosomal sequence in mammalian cells.

Replacement of the early region of simian virus 40 results in virus that cannot replicate in a normal host, CV-1 cells, but can replicate in COS cells, a derivative of CV-1 cells that constitutively express simian virus 40 tumor antigen (T antigen). However, passage of such an early replacement simian virus 40 mutant in COS cells results in the emergence of virus that can propagate in CV-1 cells. Analysis of this virus revealed that the mutant rescued the integrated T-antigen gene from the COS cell genome. Comparison of the sequence of the recovered virus with that of the viral DNA resident in COS cells (strain 776) and the mutant used in our studies (derived from strain 777) proves that the mutant virus acquired the T-antigen gene from the COS cell chromosome via homologous recombination. Most probably this process was mediated by a direct genetic exchange.

Amino Acid Sequence↗
PubMed1985

Sequence of hepatitis B virus DNA incorporated into the genome of a human hepatoma cell line.

Seven copies of integrated hepatitis B virus (HBV) DNA and contiguous genomic DNA from a human hepatoma cell line (PLC/PRF/5) have been isolated by molecular cloning and have been partially sequenced. The HBV sequences are fragmented and rearranged. Thus, the surface antigen gene is the only intact HBV transcription unit present in these integrated sequences. The sites of integration-recombination are dispersed over the entire viral genome; there is some preference for integration within the double-stranded portion of the genome. There are no repeats at the ends of the integrated HBV DNA fragments. Thus, recombination does not take place in a manner resembling the integration of retroviruses. The sequence data suggest that each HBV fragment is of the adw subtype. However, the integrated DNAs show an unexpected degree of sequence divergence. Direct evidence for the duplication, transposition, and subsequent divergence of two sequences is presented. The data surprisingly suggest that infection-integration of four distinct adw strains occurred.

Base Sequence↗
PubMed1984

Cloning and analysis of integrated hepatitis virus sequences from a human hepatoma cell line.

We report here the isolation by molecular cloning and the analysis by heteroduplex and restriction enzyme mapping of seven distinct DNA fragments containing hepatitis B virus (HBV) sequences from genomic DNA of the PLC/PRF/5 human liver carcinoma cell line (the Alexander cell). No intact full-length HBV genomes were present. Three different patterns of organization of HBV fragments were detected. These included two linear fragments without detectable rearrangement, three other HBV fragments with internal deletions, and two HBV fragments containing long inverted duplications. HBsAg sequences are preferentially included in the integrated fragment, whereas the core gene is preferentially eliminated. Several of the integrated HBV fragments might act as templates for the synthesis of functional HBsAg mRNA, whereas only one clone could produce a full core antigen transcript.

Base Sequence↗
PubMed1983

Synthesis of hepatitis B surface antigen in mammalian cells: expression of the entire gene and the coding region.

We have constructed two simian virus 40 early replacement recombinants that have the coding sequences for hepatitis B virus surface antigen (HBsAg). One construction, LSV-HBsAg, has the coding region for HBsAg but not the portion encoding the putative pre-surface antigen leader. Transformed monkey kidney cells (COS) infected with this recombinant express large quantities of the characteristic partially glycosylated HBsAg molecule, which are assembled into 22-nm particles that appear similar to those produced by human liver cells infected with hepatitis B virus. This result indicates that the pre-surface antigen sequences are not required for the synthesis of HBsAg or its assembly into particulate structures. The second recombinant, LSV-HBpresAg, has the entire surface antigen gene, including the putative promoter and pre-surface antigen region. COS cells infected with this recombinant plasmid produce 40- to 50-fold less HBsAg than those infected with the LSV-HBsAg recombinant plasmid. RNA mapping studies suggest that the transcription of the HBsAg gene is initiated at more than one site, or alternatively, that RNA splicing of transcripts occurs in the pre-surface antigen region.

Animals↗
PubMed1982

Preferential transcription and nuclear transport of globin gene sequences, as control steps leading to final differentiation of murine erythroleukemic cells.

Murine erythroleukemic (MEL) cells undergo a specific program of differentation in vitro, which is mainly characterized by accumulation of globin mRNA. These cells serve as a model system to study in detail the expression of a specific gene product at the transcriptional and post-transcriptional level. In this report we describe experiments in which the transcription rate of globin and non-globin genes, as well as their cytoplasmic appearance, was measured during the differentiation process. Two independent steps for regulating the abundance of globin mRNA were observed. On the transcriptional level we have observed that, in contrast to the transcription of globin genes, the transcription rate of non-globin genes is dramatically reduced throughout the period of induction. When the rate of cytoplasmic appearance was measured newly synthesized globin RNA molecules were found to be preferentially transported into the cytoplasm. It was shown that the reduction in cytoplasmic appearance of nonglobin genes is not a result of a shut-off in their transcriptional activity. In cells treated with 12-O-tetradecanoylphorbol 13-acetate the transcription rate remains constant while a continuous reduction in the cytoplasmic appearance is observed. These two independent phenomena which affect the non-globin genes, i.e. the suppression of their transcription and reduced cytoplasmic appearance, lead to the reduction in the relative amounts of the stable poly(A)-rich mRNA population and to the accumulation of globin sequences in the cytoplasm of the differentiated erythroid cells. These observations are in agreement with our previous model, which claimed that disappearance of the stable poly(A)-rich mRNA population is an obligatory process leading to the final differentiation of MEL cells.

Animals↗
PubMed1981

Large globin RNA molecules and their processing.

RNA containing beta-globin message sequences larger than 2000 nucleotides could be detected in nuclei of murine erythroid cells using cloned beta-globin cDNA. Under steady-state conditions, when nuclear RNA was separated on denaturing agarose gels and covalently bound to diazobenzyloxymethyl-paper, a 4200-nucleotide and a approximately equal to 3500-nucleotide band could be seen. The presence of these large molecules could also be visualized under the electron microscope after hybridization to a beta-globin genomic DNA fragment. We suggest that these molecules are precursors to mature mRNAs. In addition to these large molecules, a series of molecules smaller than 2000 nucleotides were seen. These are postulated to be processing intermediates in the maturation of beta-globin mRNA.

Animals↗
PubMed1981

Terminal direct repeats in a retrovirus-like repeated mouse gene family.

The mouse genome contains multiple copies of a dispersed gene family known individually as VL30 genes which are thought to be associated with retroviruses. The copies consist of closely related 5.2-kilobase DNA sequences flanked by unrelated cellular DNA, and although no sequence homology has been found between the genes and the exogenous or endogenous retroviruses so far tested, the 30S RNA transcripts expressed by the genes are efficiently packaged into virions, recovered from infected cells and transmitted to other cells by pseudo-type infection. Stimulated by recent reports of the similarity between retroviruses and transposons (from which retroviruses may have evolved), and in particular by the recognition that both types of genetic elements are characterized by a large terminal repeat (LTR), we set out to determine whether the VL30 genes are also distinguished by this property. Using cloned DNAs from a mouse gene library and heteroduplex analysis, we have now found that the VL30 genes do indeed carry terminal direct repeats 400 base pairs long.

Animals↗
PubMed1981

Modulation of globin gene expression by a tumor promoter during induced differentiation of Friend erythroleukemic cells.

Friend erythroleukemic cells were induced to differentiate by dimethylsulfoxide (Me2SO) in the absence or presence of the tumor promoter 12-O-tetradecanoyl-phorbol 13-acetate. The effects of the latter on the molecular parameters related to globin mRNA metabolism were examined. When differentiation was scored by benzidine staining, it had an inhibitory effect on Me2SO-treated cells. On the other hand, when differentiation was followed by determination of globin mRNA accumulation, it had a pleiotropic effect on Me2SO-treated cells. At the early phase of differentiation (2--3 days) the rate of globin mRNA accumulation was higher in the promoter-treated cells than in the control. This unexpectedly high level of accumulation was followed by a sharp reduction and most of the globin RNA sequences disappeared at later stages of differentiation (days 4--5). The reduction can be related to the effect of the promoter on the stability of globin RNA in the cytoplasm which was reduced from a half-life of 16 h to that of 8 h only. Other parameters, such as the rate of globin mRNA synthesis and its capability to serve as a template for cell-free protein synthesis were not affected by treatment with the promoter throughout the differentiation process.

Animals↗
PubMed1980

Identification of recombinant phages containing sequences from different rat myosin heavy chain genes.

The construction and identification of a recombinant plasmid containing a cDNA insert which hybridizes specifically to myosin heavy chain mRNA is described. The plasmid was used as a probe to screen a rat genomic library for recombinant phages containing myosin heavy chain sequences. Six clones with approximately 15 k bp inserts each were isolated. Digestion with several restriction enzymes and hybridization of the fractionated DNA with the plasmid probe showed that the clones contained 3 different DNA inserts. Electron microscopy of a heteroduplex made by hybridization of DNA from two clones confirmed that the inserts originated in different genes. Hybridization of size-fractionated ECOR1 digested rat spleen DNA with the cloned probe suggested the existence of at least 5 myosin heavy chain genes.

Animals↗
PubMed1977

Differences in production of solid Landhscütz tumours in BALB/c and ICR mice.

Landschütz tumour cells in the ascitic form injected subcutaneously into BALB/c or ICR mice produce solid tumours which grow progressively in most ICR mice but regress in nearly all BALB/c mice. Solid tumours in the peritoneal wall (produced by intraperitoneal inoculation of ascitic cells and treatment with normal serum) grew in both strains, but were more invasive in ICR mice. Surgical interference in BALB/c mice with these tumours, allowing adhesion of tumour to skin or subcutaneous fascia, resulted in cessation of tumour growth or regression.

Animals↗