Search PubMed⌕ Search

Biomedical subjects

E K Wagner

Publications and source records attributed to E K Wagner.

98 records · Page 6Linked to original sources
PubMed1969

RNA synthesis in cells infected with herpes simplex virus. II. Evidence that a class of viral mRNA is derived from a high molecular weight precursor synthesized in the nucleus.

Viral RNA extracted from the cytoplasmic polyribosomes of human epidermoid carcinoma no. 2 cells infected with herpes simplex virus (mRNA) had a sedimentation coefficient between 10S and 20S while that from nuclei of infected cells varied in size from 10S to >80S. Estimates of the maximum molecular weight of viral RNA from its sedimentation coefficients suggest that at least 10 per cent of the viral genome is transcribed as a single molecule. The ratio of RNA of different sizes found in the nuclei of cells pulse labeled for 12 minutes was approximately the same as those found in cells labeled for longer intervals implying that either some classes of viral mRNA were made as small molecules or that the large viral RNA molecules were cleaved soon after synthesis. Cytoplasmic mRNA competed to a level of at least 80 per cent in viral DNA-RNA hybridization tests with >50S RNA extracted from nuclei of infected cells. This is consistent with the hypothesis that viral mRNA is produced by cleavage of a large precursor RNA molecule.

Carcinoma, Squamous Cell↗
PubMed1969

Ribonucleic acid synthesis in cells infected with herpes simplex virus. I. Patterns of ribonucleic acid synthesis in productively infected cells.

HEp-2 cells were pulse-labeled at different times after infection with herpes simplex virus, and nuclear ribonucleic acid (RNA) and cytoplasmic RNA were examined. The data showed the following: (i) Analysis by acrylamide gel electrophoresis of cytoplasmic RNA of cells infected at high multiplicities [80 to 200 plaque-forming units (PFU)/cell] revealed that ribosomal RNA (rRNA) synthesis falls to less than 10% of control (uninfected cell) values by 5 hr after infection. The synthesis of 4S RNA also declined but not as rapidly, and at its lowest level it was still 20% of control values. At lower multiplicities (20 PFU), the rate of inhibition was slower than at high multiplicities. However, at all multiplicities the rates of inhibition of 18S and 28S rRNA remained identical and higher than that of 4S RNA. (ii) Analysis of nuclear RNA of cells infected at high multiplicities by sucrose density gradient centrifugation showed that the synthesis and methylation of 45S rRNA precursor continued at a reduced but significant rate (ca. 30% of control values) at times after infection when no radioactive uridine was incorporated or could be chased into 28S and 18S rRNA. This indicates that the inhibition of rRNA synthesis after herpesvirus infection is a result of two processes: a decrease in the rate of synthesis of 45S RNA and a decrease in the rate of processing of that 45S RNA that is synthesized. (iii) Hybridization of nuclear and cytoplasmic RNA of infected cells with herpesvirus DNA revealed that a significant proportion of the total viral RNA in the nucleus has a sedimentation coefficient of 50S or greater. The sedimentation coefficient of virus-specific RNA associated with cytoplasmic polyribosomes is smaller with a maximum at 16S to 20S, but there is some rapidly sedimenting RNA (> 28S) here too. (iv) Finally, there was leakage of low-molecular weight (4S) RNA from infected cells, the leakage being approximately three-fold that of uninfected cells by approximately 5 hr after infection.

Carbon Isotopes↗
PubMed1968

Effect of the Vinca alkaloids on RNA synthesis in human cells in vitro.

The Vinca alkaloids vinblastine sulfate and vincrystine sulfate, which are mitotic poisons, inhibit RNA synthesis in human (HEp-2) cells cultured in vitro. Analyses of RNA synthesis by cells treated with these drugs by acrylamide gel electrophoresis show that 28s rRNA and to a lesser extent 18s rRNA are preferentially inhibited. The synthesis of tRNA is affected much less than that of rRNA. The present experiments suggest that the drugs inhibit both the synthesis and processing of the nucleolar RNA precursors of rRNA. An explanation is also given for previous reports that these alkaloids preferentially inhibit the synthesis of tRNA in animal cells in vitro.

Antineoplastic Agents↗