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Biomedical subjects

D H Metz

Publications and source records attributed to D H Metz.

17 recordsLinked to original sources
PubMed1977

A study of events in chick cells infected with human adenovirus type 5 and their relationship to the induction of interferon.

In an attempt to define the molecular events involved in induction of interferon, various parameters of chick cells infected with human adenovirus type 5 were analysed. It was shown by digestion with various proteolytic enzymes and by disruption of the purified virus that induction of interferon requires the interaction of infectious virus with the chick cells. Analysis of adenovirus-infected chick cells by immunological and biochemical techniques indicated that most of the cells produce some virus-specific components, and that temperature sensitive mutants which fail to induce interferon at the restrictive temperature fail to synthesize late components at that temperature. However, since it has been shown that interferon can be induced in the absence of DNA synthesis, these studies conclude that interferon induction results from an early interaction between virus (or virus product) and chick cells and moreover that this interaction is also necessary for the synthesis of virus DNA in this system.

Adenoviruses, Human↗
PubMed1976

Mechanism of interferon action: further evidence for transcription as the primary site of action in simian virus 40 infection.

Interferon inhibits the replication of simian virus 40 (SV40) in monkey cells and reduces markedly the formation of both early virus protein (i.e. SV40 T antigen) and early SV40 RNA. This suggests that in SV40 infection interferon acts primarily by inhibiting transcription. To test this conclusion further, we examined alternative mechanisms which might explain these results and made the following observations. (1) The quantity of input SV40 DNA in the nucleus 24 h post infection (p.i.) was the same in interferon-treated and control cells. Thus interferon does not appear to diminish the quantity of SV40 DNA template available for transcription. (2) Chemical inhibitors of protein synthesis did not mimic the selective inhibition of early SV40 RNA formation induced by interferon, indicating that the transcription of early SV40 RNA is not dependent upon the prior synthesis of any virus-induced protein. Thus a block in translation cannot readily explain the reduced formation of early SV40 RNA in interferon-treated cells. (3) Fractionation of SV40 infected cells after a one-hour labelling period showed that interferon produced a comparable reduction in the quantity of early SV40 RNA in the nucleus and the cytoplasm. Thus the observed inhibition of early SV40 RNA is not due solely (if at all) to enhanced cytoplasmic degradation. These results indicate that the primary effect of interferon in SV40 infected monkey cells is either to inhibit the transcription of early virus RNA or to enhance its turnover in the nucleus.

Cell Line↗
PubMed1976

Further investigations on the mode of entry of vaccinia virus into cells.

Earlier results indicating that vaccinia virus entered L cells by a process of direct fusion between the virus envelope and the plasma membrane of the cell have been confirmed and extended using immuno-ferritin conjugates to locate virus antigens on the host cell surface. After fusion, components of the virus envelope become rapidly dispersed in the plasma membrane. Fusion has also been observed as the predominant mode of entry of vaccinia virus into HeLa cells.

Adsorption↗
PubMed1975

The formation of virus polyribosomes in L cells infected with vaccinia virus.

The fate of early virus messenger RNA in the cytoplasm of vaccinia-infected L cells has been studied during the first hour after infection. The RNA is made in the virus core structure from which it is rapidly released. It accumulates in the polyribsome fraction, where at least 75% is bound to ribosomes through an EDTA-sensitive link. Three distinct structures have been identified as possible intermediates in virus polyribosome formation. The first is a ribonucleoprotein complex (RNP) in which virus RNA is associated with cellular proteins. A complex having apparently similar properties, is formed when virus RNA is added to a cytoplasmic extract in vitro. The other two structures may consist of an RNP moiety associated with the small ribosomal subunit, or with a single ribosome. At least part of the RNA isolated as RNP appears to be a precursor of the virus messenger found in polyribosomes.

Animals↗
PubMed1975

The effect of interferon on the formation of virus polyribosomes in L cells infected with vaccinia virus.

The effect of interferon treatment of mouse L cells on the fate of virus messenger RNA following infection with vaccinia virus has been studied. The polyribosomes of interferon-treated, infected cells are found to be disaggregated and it is proposed that htis results from inhibition of the initiation of virus polypeptide snythesis. Evidence is presented that inhibition of polypeptide chain elongation also occurs. The block in initiation appears to be due to the failure of the small ribosome subunit to attach to the virus messenger ribonucleoprotein complex. The translation of the different vaccinia messenger species is inhibited to a comparable extent.

Animals↗
PubMed1972

Mechanism of interferon action: inhibition of viral messenger ribonucleic acid translation in L-cell extracts.

Encephalomyocarditis (EMC) virus ribonucleic acid (RNA) stimulated the incorporation of (14)C-amino acids into polypeptides in cell-free systems using preincubated S10 extracts from L cells. Incorporation was linear for over 2 hr. Analysis of the tryptic peptides derived from the polypeptide products formed in response to EMC RNA showed them to be virus specific. The major product, a polypeptide of 140,000 in molecular weight, migrated on sodium dodecyl sulfate-polyacrylamide gels with one of the virus-specific polypeptides present in EMC-infected cells. A minor component of molecular weight about 230,000 may correspond to the product of complete translation of the EMC virus genome. Little or no effect of interferon or vaccinia virus infection was observed in the preincubated, cell-free system. The EMC RNA-stimulated incorporation of (14)C-amino acids into polypeptides was not inhibited in extracts derived from L cells early in virus infection, from interferon-treated cells, or from cells subjected to both treatments. Interferon treatment did appear to have a slight inhibitory effect on chain elongation in this system. However, treatment of cells with highly purified interferon before virus infection caused a decrease of about 80% in the capacity of non-preincubated cell extracts to translate added EMC RNA. This effect did not extend to the translation of polyuridylic acid and could be reversed by preincubation of the extracts at 37 C for 20 min. The inhibition of translation was manifest at interferon concentrations as low as 5IU/ml, and in this respect closely paralleled the inhibition of virus growth. Inactivation of the antiviral activity of the interferon by heating or digestion with trypsin also abolished the effect on cell-free protein synthesis. The EMC-specific polypeptides formed in reduced amounts in extracts of interferon-treated vaccinia-infected cells were smaller than those formed in extracts of untreated, vaccinia-infected cells. Thus, inhibition of initiation or elongation of polypeptides, or both, can be demonstrated in cell-free systems employing non-preincubated extracts from interferon-treated, virus-infected cells. These results indicate that antiviral activity of interferon is directed against the translation of viral messenger RNA.

Amino Acids↗