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M J Sekellick

Publications and source records attributed to M J Sekellick.

At least 37 records · Page 2Linked to original sources
PubMed1989

Interferon induction by viruses. XVII. Non-temperature-sensitive mutations regulate interferon induction by vesicular stomatitis virus.

Wild-type (wt) strains of vesicular stomatitis virus (VSV) strain Indiana are poor to non-inducers of interferon (IFN) which express IFN induction-suppressing activity. At non-permissive temperatures, temperature-sensitive (ts) mutants of this virus are either like their wt parents, or they are good to excellent inducers of IFN. IFN inducibility and IFN induction-suppressing activity are mutually exclusive phenotypes in VSV-Indiana. With one exception, all Orsay ts mutants derived by A. Flamand (CNRS, Gif-sur-Yvette, France), representing the five complementation groups, were poor to non-inducers of IFN and were also capable of suppressing IFN induction by other viruses. In contrast, all Glasgow ts mutants derived by C. R. Pringle (University of Warwick, Coventry, U.K.) were excellent inducers of IFN. We demonstrate that this difference in acquisition of IFN inducibility relates primarily to the origin of the mutations; spontaneous for Orsay, and mutagen-derived for Glasgow. Tests with newly generated spontaneous and mutagen-derived mutants, and temperature-stable revertants of IFN-inducing ts mutants indicate that IFN inducibility results from non-ts, multiple mutations rarely acquired spontaneously, but generated frequently upon mutagenesis with 5-fluorouracil. The capacity of VSV-Indiana to induce IFN is considered intrinsic to the virus, but is only manifested when the dominant IFN induction-suppressing phenotype is lost through mutagenesis. Thus, non-ts mutations appear to regulate the expression of the IFN induction-suppressing phenotype, and hence the IFN inducibility of VSV-Indiana.

Animals↗
PubMed1988

Interferon induction by viruses. XVI. 2-Aminopurine blocks selectively and reversibly an early stage in interferon induction.

A purine analogue, 2-aminopurine, reported to act as an inhibitor of protein kinase, selectively, reversibly and in a dose-dependent manner blocked a very early stage in interferon induction. With chick embryo cells and mouse L cells as hosts, and different viral inducers of interferon, maximal effects of 2-aminopurine were observed during the first 4 h of induction. At 10 mM-2-aminopurine there was a 20-fold reduction in the yield of interferon from both cell types. 2-Aminopurine and actinomycin D both prevented interferon induction with the same time course, indicating a transcriptional block to induction; however, only the action of the former was reversed upon removal of the drug. Addition of 2-aminopurine to an agarose overlay resulted in high efficiency plaque formation by vesicular stomatitis virus New Jersey (Hazelhurst) under conditions where endogenous induction of interferon and its feedback action on aged chick embryo cells normally prevented plaque formation. Two other inducible systems, representing genes involved in interferon action (both its development and activation), and those of heat shock, were not affected by 2-aminopurine. A model is presented implicating the interferon-inducible dsRNA-dependent protein kinase as an interferon induction receptor which, on interaction with dsRNA, generates an amplified signal via phosphorylation that ultimately derepresses the interferon gene(s).

2-Aminopurine↗
PubMed1987

Interferon induction by viruses. XV. Biological characteristics of interferon induction-suppressing particles of vesicular stomatitis virus.

A single interferon (IFN) induction-suppressing particle (ISP) of vesicular stomatitis virus (VSV) blocked completely the yield of IFN in a cell otherwise programmed to produce IFN. With mouse L cells as hosts, one lethal hit of UV radiation (D37 = 52.5 ergs/mm2) to the VSV genome sufficed to inactivate ISP activity; however, with "aged" primary chick embryo cells as hosts, it took 198 lethal hits (D37 = 10,395 ergs/mm2). ISP expression in chick cells did not require virus replication or amplified RNA synthesis, but did involve functional virion-associated L protein. ISP in chick cells also were capable of inhibiting, in a multiplicity-dependent manner, the plaquing efficiency of two viruses that require cellular polymerase II (pol II) for replication, e.g., pseudorabies and influenza. The refractory state to IFN inducibility that resulted from infection of chick cells with ISP (VSV tsO5 [UV = 100 hits]) was still extant after 6 days. In contrast, the plaquing efficiency of pseudorabies virus returned to control levels by 5 h after ISP infection. Chick cells infected with UV ISP remained viable, served as hosts for the replication of other viruses, and could be subcultured. Models are presented to account for these contrasting effects. The involvement of viral plus-strand leader RNA as an inhibitor of cellular pol II-dependent RNA synthesis, and the multifunctional activities of the virion-associated L protein, are discussed as possible molecules involved in the action of ISP in chick cells.

Animals↗
PubMed1985

Interferon induction by viruses. XIII. Detection and assay of interferon induction-suppressing particles.

An assay is described for detecting and measuring the capacity of a single virus particle to suppress completely the yield of interferon from a cell otherwise competent to product interferon. The assay defines virus populations in terms of a newly quantifiable attribute--interferon induction-suppressing particles (ISP). The antagonistic actions of interferon-inducing particles and interferon induction-suppressing particles are thought to determine the interferon-inducing capacity intrinsic to a virus population.

Animals↗
PubMed1985

Interferon induction by viruses. XIV. Development of interferon inducibility and its inhibition in chick embryo cells "aged" in vitro.

Studies with a number of viruses revealed a time-dependent acquisition of interferon (IFN) inducibility in primary chick embryo cells as they "aged" in vitro for 2-12 days at a confluent cell density without a medium change. The time-course for the development of IFN inducibility was established by generating and analyzing a family of dose (multiplicity)-response (IFN yield) curves, using Newcastle disease virus (NDV, strain LaSota) as the inducer. Cells produced little or no IFN for the first 4-5 days. Between 5 and 6 days the cells gradually developed the capacity to respond to NDV (and other viruses). Maximal yields of IFN were inducible by day 10. This time-dependent development of IFN inducibility was abrogated almost completely when "aging" was carried out in the presence of drugs that inhibited the synthesis of cyclic derivatives of C20 oxygenated unsaturated fatty acids, i.e., inhibitors of prostaglandin/leukotriene synthesis and the arachidonic acid cascade. Of the prostaglandin synthesis inhibitors, indomethacin was particularly effective. Cells treated on day 0 with 10 micrograms/ml of indomethacin produced 100- to 1000-fold less IFN than controls when induced on day 8. To prevent maximally the development of IFN inducibility, indomethacin must be added within the first 2 days of seeding. After about 2 days, the cells begin to escape the action of the drug. Indomethacin added at the time of induction had no effect on the yield of IFN. IFN inducibility was partially restored when indomethacin was removed during the aging process. "Aging" chick cells in low concentrations of cycloheximide (0.5 micrograms/ml) produced results comparable to incubation with indomethacin. Neither reagent had any marked effect on the rate of total protein or RNA synthesis, nor did their action prevent the induction of stress (heat shock) proteins. Cells "aged" in indomethacin were threefold less efficient in responding to the action of IFN, whereas aging in cycloheximide had no effect on IFN's action. Plaque formation on drug-treated cells was normal for viruses that were poor inducers of IFN. However, both the plaquing efficiency and plaque size of Sindbis virus (an excellent IFN inducer) were enhanced markedly on cells treated with indomethacin or low concentrations of cycloheximide during the aging process. These data implicate a family of fatty acid derivatives of arachadonic acid, including prostaglandins and leukotrienes, in the development of IFN inducibility in primary chick embryo cells "aged" in vitro.

Animals↗
PubMed1983

Interferon induction by viruses. X. A model for interferon induction by Newcastle disease virus.

Unirradiated Newcastle disease virus (NDV, strain AV) induced high levels of interferon (IFN) in primary chick embryo cells if the cells were 'aged' in vitro for 6 to 7 days. Dose (multiplicity)-response (IFN yield) curves, carried out in the presence of anti-NDV serum to prevent cycling infection, revealed that stocks of NDV-AV contain about sevenfold more IFN-inducing particles (IFP) than infectious particles (PFP). These non-infectious IFP were responsible for nearly all IFN induction in 'aged' cells, since PFP were determined to be incapable of inducing IFN. In contrast, with mouse L(Y) cells as hosts, about one-third the number of particles as there are PFP appeared to score as IFP. Heat and u.v. radiation (254 nm) inactivated NDV IFP and PFP activity at the same rate whether tested in chick or mouse cells, implying that virion-associated transcription is required to induce IFN. A model is proposed to account for the generation of IFN-inducing particles from infectious NDV following u.v. irradiation, and their subsequent inactivation at high doses of radiation. The model defines a series of u.v. targets in the NDV genome that regulate the expression of IFN-inducing particle activity in 'unaged' chick embryo cells.

Animals↗
PubMed1980

Interferon induction by viruses. III. Vesicular stomatitis virus: interferon-inducing particle activity requires partial transcription of gene N.

We have measured the interferon-inducing particle (i.f.p.) activity of a ts mutant, GII (I), of vesicular stomatitis virus (VSV) and a non-ts revertant, RI (T1026) in "aged" chick embryo cells and mouse (L(Y) cells at 40.5 and 37.5 degrees C, respectively. Our results suggest that a single i.f.p. suffices to induce a quantum yield of interferon and that there are several times more i.f.p. than plaque-forming particles (p.f.p.) in stock preparations of VSV. Furthermore, while virus replication or amplified RNA synthesis is not required for a particle of VSV to induce interferon, there is a requirement for primary transcription. About one-tenth of the genome must remain intact and be transcribed to synthesize an interferon-inducer moiety. (This represents transcription of about two-thirds of the N protein gene.) We conclude that VSV does not contain a pre-formed inducer of interferon and propose a model for its formation. We suggest that there is a cumulative loss of N (and/or NS and L) protein from the ribonucleoprotein complex during primary transcription, leading ultimately to extensive base-pairing between the genome RNA and its complementary transcript. We suggest that the dsRNA thus formed constitutes the interferon inducer moiety of VSV.

Animals↗
PubMed1978

Interferon action III. The rate of primary transcription of vesicular stomatitis virus is inhibited by interferon action.

Transcription of vesicular stomatitis virus (VSV) in Vero cells was confined to the synthesis of parentally-derived mRNA (primary transcription) by the use of cycloheximide and/or a ts mutant, G41(IV), at a non-permissive temperature (40 degrees C). More transcripts accumulated in the presence of cycloheximide than in its absence. This so-called "cycloheximide effect" results from higher rates of virus transcription sustained for longer periods of time. The rate of VSV transcription initially increases linearly for 1 to 2 h after injection. Interferon reduces this rate (congruent to fourfold with 50 units/ml interferon) irrespective of the presence or absence of cycloheximide. The VSV mRNA transcripts synthesized in mock- or interferon-treated cells were equal in size and had an equivalent half-life of 17 h at 40 degrees C. It seems likely that once transcription is initiated in interferon-treated cells, it is completed successfully. Since interferon reduces the rate of early VSV primary transcript synthesis to below that achieved in the presence of cycloheximide, we conclude that interferon has an effect on transcription beyond that attributable solely to protein synthesis inhibition. We postulate that interferon decreases the probabiltiy of initiating virus transcription. Virus mRNA escaping this facet of interferon action may then encounter other facets such as post-transcriptional modification and/or inhibition of translation. However, the mandatory sequence of primary transcription leads to primary translation for negative-strand viruses like VSV dictates that the overall inhibitory effect of interferon on translation would derive in part from this prior inhibition of transcription. Thus, to apply the term "primary effect" to one particular facet of interferon action may not always be meaningful.

Cycloheximide↗