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Biomedical subjects

G Winberg

Publications and source records attributed to G Winberg.

45 records · Page 3Linked to original sources

Expression of the Epstein-Barr virus encoded EBNA-1 gene in stably transfected human and murine cell lines.

Five murine and 3 human tumor cell lines were transfected with a retroviral vector that carries the EBV encoded EBNA-1 gene. All cell lines expressed intranuclear EBNA-1 as detected by anticomplement immunofluorescence and Western blot assays. The cell lines differed in the level of EBNA-1 expression and the size of the protein. The internal major late promoter of adenovirus was efficient in directing the transcription of EBNA-1 in the human lymphoma line BJAB, the murine T-cell lymphoma Tikaut, RBL-5, EL-4 and in the mouse sarcoma line MSWBS but was less efficient in Ramos, an EBV negative Burkitt lymphoma line, the human T-cell leukemia line 1301TK and the P815-X2 mouse mastocytoma line. All transfected lines except MSWBS contained EBNA-1 in a truncated form. The truncated EBNA-1 polypeptide reacted with the conventional human antibody reagents in an EBNA specific fashion but failed to bind rabbit or human antibody directed against the glycine-alanine repeat sequence. MSWBS contained a truncated as well as a full size EBNA-1 polypeptide. It also reacted with antibody directed against the glycine-alanine repeat. This indicates that the repeat sequence is regularly affected by the truncation.

Adenoviridae↗

Infection of a murine T-cell line with a retrovirus carrying c-myc decreases the interleukin-2 cell dependence by a nonautocrine mechanism.

A murine interleukin-2 (IL-2)-dependent T-cell line, CTLL-2, was infected with a retrovirus carrying the mouse c-myc cDNA and the Tn-5 neogene. Transduced cells were selected in the presence of Geneticin sulfate (G418); these cells were shown to express both the endogenous and the transduced c-myc genes. The IL-2 requirement of these cells was then found to be significantly reduced. The cells did not express IL-2 mRNA nor did they produce an activity mitogenic for CTLL-2 cells. This suggests that the reduction of IL-2 dependence observed following retroviral transduction of c-myc is caused by a nonautocrine mechanism.

Animals↗

Dissection of overlapping functions within the adenovirus type 5 E1A gene.

The adenovirus E1A gene encodes multiple, overlapping mRNAs whose products function both to regulate mRNA levels during the lytic cycle of the virus and to facilitate transformation of non-permissive cells. To assign specific functions to the E1A gene products, two adenovirus type 5 variants have been constructed. Mutants dl347 and 348 carry cloned segments corresponding to the E1A 12 and 13S mRNAs, respectively, in place of the normal E1A gene. The variants produced the predicted E1A-specific mRNAs and polypeptides. Both viruses grew efficiently in HeLa cells. Although the 13S mRNA products were more effective, the products of either mRNA species could stimulate the accumulation of mRNAs from additional transcription units. Both viruses could induce the formation of transformed foci in an established rat cell line. Neither virus could transform primary rat embryo cells at normal frequency, and the dl347 foci which were induced were incomplete or abortive transformants. Thus, functions encoded by both 12S and 13S mRNAs are required for efficient and complete transformation of primary rat cells.

Adenoviridae↗

Isolation of DNA from agarose gels using DEAE-paper. Application to restriction site mapping of adenovirus type 16 DNA.

A new method for isolating DNA from agarose gels is described. The method involves the simultaneous transfer of all DNA-fragments from an agarose slab gel onto DEAE-cellulose paper and the elution of the individual fragments from the paper with 1 M NaCl. DNA isolated from agarose gels in this way is susceptible to cleavage with several restriction endonucleases, and can be labeled in vitro with E coli DNA-polymerase I, T4 DNA-polymerase and T4 polynucleotide kinase. We have used the method to construct restriction endonuclease maps of adenovirus type 16 DNA.

Adenoviridae↗

Structural polypeptides of adenovirus type 16 incomplete particles.

The polypeptides of adenovirus type 16 incomplete particles, with average buoyant densities of CsCl of 1.33, 1.318, 1.305, and 1.299 g/cm3 and DNA content of less than 1 U genome size, were compared with the polypeptides of the complete virion (density, 1.344 g/cm3, and 22 x 10(6) daltons of DNA) and late polypeptides in infected cells by using sodium dodecyl sulfate-gel electrophoresis. In agreement with other serotypes studied (types 2 and 3), the light particles lack polypeptides V, VI, and VII. In adenovirus type 16, eight other major polypeptides are found, with apparent molecular weights of 59,000 (59K), 46K, 31K, 30K, 28K, 27K, 26K, and 19K. The 30K to 31K and 27K to 28K polypeptides are phosphorylated. The 27K and 19K polypeptides are precursors, whereas the 31K, 30K, and 26K polypeptides are chase products in the cell, as are polypeptides VI, VII, and VIII. The 26K polypeptide is proposed to be an intermediate in processing since it disappears from the young virions upon chasing. Although chase products, the 31K and 30K polypeptides are only associated with particles having buoyant density lower than 1.318 g/cm3. Polypeptides V and VII are only present in particles containing more than one quarter of the genome. No trace of cell histones could be detected in the purified incomplete particles. A major constituent of the incomplete particles, the 46K polypeptide, was rapidly labeled in the cell, and loss of radioactivity from this band was detectable only after 7 to 18 h of chase.

Adenoviruses, Human↗