bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic diversity in natural populations of mammalian reoviruses: tryptic peptide analysis of outer capsid polypeptides of murine, bovine, and human type 1 and 3 reovirus strains
Authors: Gentsch J R , Fields B N
Published: January 2020

We have studied the structural relationships between the outer capsid polypeptides of eight murine, bovine, and human isolates of type 1 and 3 mammalian reoviruses. Our results show that the outer capsid polypeptides of reoviruses isolated from different mammalian species, in different years and different geographical areas, have both conserved and unique methionine-containing tryptic peptides. We found that tryptic peptides from mu 1C polypeptides of two human, one murine, and two bovine type 3 isolates and one human and two bovine type 1 reoviruses are highly conserved. Our data show that only one tryptic peptide pattern of the mu 1C polypeptide (encoded by the M2 gene) was present in reoviruses isolated from the three different mammalian species. The mu 1C polypeptide of the type 3 Dearing strain contained one tryptic peptide not found in any other reovirus isolate examined. In marked contrast to the mu 1C polypeptides, the sigma 3 polypeptides (encoded by the S4 gene) of three type 1 and three type 3 isolates were divided into two patterns based on significant differences in their tryptic peptides. In addition, at least seven tryptic peptides were conserved among the sigma 3 polypeptides of all virus strains examined. The sigma 3 polypeptide of the type 3 Dearing strain was distinguishable from the sigma 3 polypeptides of all other strains examined. The one mu 1C and two sigma 3 tryptic peptide patterns were found to occur interchangeably in isolates of type 1 or type 3. About 1/3 of the tyrosine-containing tryptic peptides of sigma 1 polypeptides of four type 3 isolates examined were conserved. Comparison of peptide differences in sigma 1 polypeptides of these isolates showed that each had one or more unique tryptic peptides, suggesting that the S1 genes coding for these polypeptides had undergone genetic drift or, alternatively, that there are at least two tryptic peptide patterns present among the sigma 1 polypeptides of these isolates. Our results suggest that genetic drift and reassortment are the most likely explanation for the extensive genetic diversity found in natural populations of mammalian reoviruses.
Characterization of La Crosse virus small-genome transcripts
Authors: Patterson J L , Kolakofsky D
Published: January 2020

With restriction fragments from DNA clones of the La Crosse virus S genome segment, the 3' end of the S mRNA was located by S1 nuclease mapping near a polyuridine tract, approximately 100 nucleotides, from the end of the S genome. Genome replication in La Crosse virus-infected cells was abolished by the drug cycloheximide, similar to other negative-strand RNA viruses. However, the synthesis of S mRNA could not be detected in cells pretreated with cycloheximide, suggesting that ongoing protein synthesis is required for La Crosse virus genome transcription and replication. Primer extension experiments in the presence of chain-terminating nucleoside triphosphates demonstrated that the 5' end of the La Crosse virus S mRNA begins 10 to 14 nucleotides before the 3' end of the S genome segment, suggesting that the La Crosse virus S mRNA is initiated on a host primer. A hypothesis consistent with these unexpected findings is presented.
A transcript from the S segment of the Germiston bunyavirus is uncapped and codes for the nucleoprotein and a nonstructural protein
Authors: Bouloy M , Vialat P , Girard M , Pardigon N
Published: January 2020

Analysis of the RNAs present in BHK-21 cells infected with Germiston virus showed that the transcripts from the L and M segments have a size similar to that of their template, whereas two types of complementary RNA are transcribed from the S segment. One, S1, is a full-length "plus" RNA strand (antigenome), and the other, S2, is an incomplete plus RNA strand which serves as mRNA for at least the synthesis of the N protein and a virus-specific nonstructural polypeptide, p12. The 5' ends of these two transcripts appeared to be identical and complementary to the 3' ends of the viral RNA. Our results suggest that transcription of the S fragment either stops 100 to 150 nucleotides from the 5' end of the template, generating an S2 molecule, or continues, generating an S1 molecule. Neither the S1 antigenome nor the S2 mRNA molecules were polyadenylated at their 3' ends or capped at their 5' ends.
Organization of the left-hand end of the herpes simplex virus type 2 BglII N fragment
Authors: Galloway D A , Swain M A
Published: January 2020

We have determined the complete nucleotide sequence surrounding the region coding for a 38,000-dalton protein of herpes simplex virus type 2 strain 333 that is encoded between map coordinates 0.58 to 0.595 on the viral genome. The sequence data have revealed an open translational reading frame of 1,011 nucleotides encoding a protein of 337 amino acids. Upstream of those sequences, transcriptional regulatory signals could be identified that overlap another open reading frame of 489 nucleotides, which presumably encodes the carboxy-terminal 163 amino acids of a 140,000-dalton protein. Downstream of the gene encoding the 38,000-dalton protein a polyadenylation signal was found as well as one further along on the other strand near a termination codon that presumably punctuates the gene specifying a 61,000-dalton protein. The DNA sequence data were compared with the amended sequence of the herpes simplex virus type 1 strain KOS 40,000-dalton protein (K. G. Draper, R. F. Frink, and E. K. Wagner, J. Virol. 43:1123-1128, 1982) and with the 5' end of the same gene from herpes simplex virus type 1 strain 17 (J. McLaughlan and J. B. Clements, J. Gen. Virol. 64:997-1006, 1983) to assess the degree of inter- and intratypic variation. The comparison of the sequences has provided a basis for the type specifity of this viral protein and for different mechanisms giving rise to the diversity between herpes simplex virus types 1 and 2.
Characterization of viable mutants of polyomavirus cold sensitive for maintenance of cell transformation
Authors: Templeton D , Eckhart W
Published: January 2020

We mutagenized a cloned fragment of polyoma DNA encoding portions of the middle size (MT) and large T antigens. We regenerated infectious viral genomes containing the mutagenized DNA and tested their transforming ability at 32 and 39 degrees C. We isolated three nontransforming mutants and two mutants which were cold sensitive for the maintenance of cell transformation. The nontransforming mutants contained amber termination codons in the reading frame for the MT antigen. They synthesized truncated MT antigens which lacked MT-associated protein kinase activity. The cold-sensitive mutants synthesized MT antigens indistinguishable from wild type with regard to size, stability at 32 and 39 degrees C, intracellular location, and associated protein kinase activity. One of the mutants was shown by nucleotide sequence analysis to contain a single amino acid change in the MT antigen, located two residues upstream from the C-terminal hydrophobic region, and no changes in the large T antigen. The other mutant contained two amino acid changes in the MT antigen and two amino acid changes in the large T antigen.
Increased concentration of an indigenous proviral mouse mammary tumor virus long terminal repeat-containing transcript is associated with neoplastic transformation of mammary epithelium in C3H/Sm mice
Authors: Graham D E , Medina D , Smith G H
Published: January 2020

Increased amounts of mouse mammary tumor virus (MMTV) proviral transcripts were found in RNA dot blots from MMTV-negative, C3H/Sm mouse mammary tumors which arose spontaneously or were induced by hormonal or chemical carcinogens or both. Other dot blots probed with a long terminal repeat (LTR) probe showed that LTR (MMTV)-containing transcripts were disproportionately represented in these tumor RNAs. Different segments of the MMTV genome were used in sequential hybridizations to Northern blots to determine relative sequence content and size of MMTV transcripts in transformed mammary tissues, as compared with those in lactating mammary glands. Increased amounts of 4.4-kilobase env and 8.1-kilobase genomic MMTV transcripts were detected with an env probe in many of the tumor RNAs examined. Hybridization of the same Northern blots containing tumor RNAs with an LTR probe revealed a 2.2-kilobase transcript which was prominent in RNAs from chemically-induced, hormonally-induced, and spontaneous mammary tumors relative to those from lactating mammary glands. The LTR-containing transcript did not possess significant homology to either env or gag-pol probes. This distinctive, transformation-enhanced, 2.2-kilobase transcript may contain mouse cellular sequences in addition to LTR sequences or it may represent the message for a nonstructural viral protein encoded within the LTR open reading frame of one or more of the four C3H/Sm MMTV proviral genes.
Target cells for avian myeloblastosis virus in embryonic yolk sac and relationship of cell differentiation to cell transformation
Authors: Boettiger D , Durban E
Published: January 2020

The yolk sac of the 12-day chicken embryo retains the blast stage progenitors to cells of the myeloid lineages with a very low level of contamination by more mature myeloid cells which have begun to express the characteristic myeloid cell markers. Both in vivo and in vitro experiments have supported the hypothesis that target cells for the BAI-A strain of avian myeloblastosis virus are contained within the myeloid lineages. An assay system for avian myeloblastosis virus was developed which utilizes this yolk sac cell system and which appears to be more sensitive than previous published assays. In addition, the kinetics of a liquid culture transformation system is presented in which at least 4% of the yolk sac cell population was transformed in a relatively synchronous fashion at 2 days after infection. The morphological transformation preceded an increased rate of cell proliferation. Cell separation procedures provided a 10- to 20-fold enrichment of target cells and demonstrated that the target cell population copurifies with macrophage colony-forming cells which are the committed progenitors to the macrophage lineage. In combination with earlier work, this work demonstrated that cells committed to the macrophage lineage at all stages of differentiation may serve as target cells for infection by avian myeloblastosis virus.
Protein processing map of poliovirus
Authors: Pallansch M A , Kew O M , Semler B L , Omilianowski D R , Anderson C W , Wimmer E , Rueckert R R
Published: January 2020

Five previously unmapped proteins (5a, 7d, 8, 9b, and 10) were located on the proteolytic processing map of the polyprotein. One of the proteins, 9b, appears to be the sister fragment of a cleavage reaction (P3-9 leads to P3-9b + VPg). Two of the other newly mapped proteins, 8 and 10, have been identified as sister fragments of X-related proteins 3b and 5b; thus, P2-3b leads to P2-8 + P2-5b and P2-5b leads to P2-10 + P2-X. The remaining proteins, 5a and 7d, mapped in the 1b protein and appear to result from the cleavages P3-1b leads to P3-5a + P3-6b and P3-4b leads to P3-7d + P3-6b. These assignments account for over 95% of the total polioviral proteins and complete the mapping of the major processing pathways.
Polyomavirus and simian virus 40 large T antigens bind to common DNA sequences
Authors: Pomerantz B J , Hassell J A
Published: January 2020

The large T antigens of polyomavirus and simian virus 40 (SV40) recognize and bind to specific, noncoding DNA sequences which are located between the beginning of the early and late transcription units in their respective genomes. Each large T antigen binds to multiple sites within this intergenic DNA stretch. Polyomavirus large T antigen binds to at least two sites within its DNA, and SV40 large T antigen binds to three sites within SV40 DNA. Comparison of the DNA sequences which comprise the binding sites in polyomavirus DNA or those which make up the binding sites in SV40 DNA has led to recognition of a common sequence, -GAGGC-, which is repeated within each large-T-antigen-binding site. We tested the hypothesis that repeats of this pentanucleotide form the recognition-binding site for polyomavirus and SV40 large T antigen. This was accomplished by measuring the binding of each large T antigen to both polyomavirus and SV40 DNA and to synthetic DNA substrates which did or did not contain repeats of the -GAGGC- sequence. Polyomavirus large T antigen bound to specific fragments of SV40 DNA, and SV40 large T antigen bound with specificity to polyomavirus DNA. In each case, the DNA fragments bound by the heterologous large T antigen were the same as those bound by the homologous large T antigen. Moreover, polyomavirus and SV40 large T antigen only bound to synthetic DNA substrates which contained repeats of the pentameric sequence. This synthetic DNA also competed effectively with native polyomavirus or SV40 DNA as a substrate in binding reactions with one or the other large T antigen. These results led us to conclude that repeats of the -GAGGC- sequence form the recognition-binding site for both polyomavirus and SV40 large T antigen.
Herpesvirus saimiri DNA in a lymphoid cell line established by in vitro transformation
Authors: Schirm S , Müller I , Desrosiers R C , Fleckenstein B
Published: January 2020

A lymphoid T-cell line (H1591) was established by infecting peripheral blood mononuclear cells from a cotton top marmoset with Herpesvirus saimiri OMI. Analysis of these in vitro-immortalized cells revealed nonintegrated, covalently closed circular viral DNA molecules in high multiplicities with substantial rearrangements and large deletions in their L-DNA (unique) regions. One subline, designated H1591 Er, contained circular viral DNA with one stretch of H-DNA (repetitive) and one of L-DNA; the L-DNA segment consisted of a linear fusion of a 53.2-kilobase-pair piece of L-DNA (left half of L-DNA) with a 15.2-kilobase-pair L-DNA fragment from the right end of the L-DNA region. The other subline, H1591 S, contained two short regions of L-DNA, each derived from the extreme ends of virion L-DNA. Both L-DNA regions of H1591 S cells contained inverted repetitions (15.0 +/- 0.2 and 9.1 +/- 4.7 kilobase pairs). The extensive deletions of L-DNA sequences in cell line H1591 indicate that at least 73% of the genetic information in H. saimiri is not required to maintain the persistence of viral DNA and the state of transformation in lymphoid T-cells.
Deletions in the genomes of pseudorabies virus vaccine strains and existence of four isomers of the genomes
Authors: Lomniczi B , Blankenship M L , Ben-Porat T
Published: January 2020

As part of a study designed to identify the genes responsible for the virulence of pseudorabies virus, we have mapped the genomes of two independently derived vaccine strains (Bartha and Norden) by restriction enzyme analysis. The structures of these genomes have been compared with that of the genome of a laboratory strain previously mapped, of restriction fragments which had been cloned. The genome of the Bartha strain was found to be very similar to that of other pseudorabies virus strains, except that a deletion of approximately 2.7 X 10(6) daltons was found in the unique short (US) region. This deletion was also observed in the genome of the Norden vaccine strain but was not observed in the genomes of any other pseudorabies virus strains that have been studied (more than 20). The genome of the Norden strain differs from that of other pseudorabies virus strains in several other respects as well. The most important difference is that in contrast to all other pseudorabies virus strains analyzed to date, which contain a type 2 herpesvirus DNA molecule (in which the US region only inverts itself relative to the unique long [UL] region), the genome of the Norden strain is a type 3 molecule in which both the US and the UL regions of the genome invert themselves, giving rise to four isomeric forms of the genome. The ability of the UL region to invert itself is probably related to the fact that a sequence normally present in all other pseudorabies virus strains at the end of the UL only is found also in inverted form at the junction of the UL and the internal inverted repeat in the Norden strain.
Glycoprotein gp118 of varicella-zoster virus: purification by serial affinity chromatography
Authors: Friedrichs W E , Grose C
Published: January 2020

Glycoprotein gp118, one of the major glycosylated proteins specified by varicella-zoster virus, is biologically of great importance since it possesses an epitope which elicits a complement-independent neutralizing antibody response. To purify this glycoprotein from a Nonidet-solubilized extract of varicella-zoster virus-infected cells, we examined its affinity to a variety of ligands, including two lectins--concanavalin A and Lens culinaris, Cibacron blue and heparin, and finally an immunoadsorbent anti-gp118 monoclonal antibody. By serial affinity chromatography on three different columns consisting of, respectively (i) Cibacron blue dye-Sepharose, (ii) L. culinaris-Sepharose, and (iii) anti-gp118 murine monoclonal antibody bound to CNBr-activated Sepharose, we isolated varicella-zoster virus-specific gp118 essentially free of contamination by any other radiolabeled viral or cellular polypeptide. The fold purification was estimated at 1,025 and the percent recovery at 13.6. On the basis of its chromatographic properties, gp118 appeared to contain mainly asparagine-linked, biantennary, complex-type, and hybrid-type oligosaccharides.
Multiplication of Bacteriophage P22 in Penicillin-Induced Spheroplasts of
Salmonella typhimurium
Authors: Ray R. K. , Burma D. P.
Published: January 2020

The spheroplasts of
Salmonella typhimurium
(LT2) prepared by treatment with penicillin were capable of adsorbing phage P22 C
1
. The normal multiplication of the phage took place, although the burst size was reduced to one-fourth of that in intact cells. Rate of incorporation of
14
C-thymidine into spheroplasts was increased severalfold on phage infection. Multiplication of C
+
also took place, but no lysogeny could be established in spheroplasts. Furthermore, spheroplasts prepared from cells lysogenized with wild-type phage, LT2 (C
+
), and a temperature-inducible C
2
mutant, LT2(tsC
2
), were not inducible. Unlike normal cells, both mitomycin C and actinomycin D interfered with the phage multiplication in spheroplasts. The spheroplast system offers great advantages in the study of the synthesis of nucleic acids and proteins in phage-infected LT2.
Control of Gene Function in Bacteriophage T4
Authors: Sauerbier Walter , Bräutigam Alan R.
Published: January 2020

Replication of T4 deoxyribonucleic acid (DNA) is known to be required for the onset of transcription of late T4 genes. Once late gene transcription has been initiated, DNA replication is no longer required for maintaining synthesis of late or early T4 messenger ribonucleic acid (mRNA). Late phage proteins (lysozyme and tail fibers) continue to be produced at constant rates after interrupting T4 DNA synthesis. The ability of the host cell to glucosylate the T4 progeny DNA has no demonstrable influence on the rates at which T4 mRNA and late proteins are synthesized after the interruption of DNA synthesis. To explain the requirement of T4 DNA replication for the onset of late gene transcription, we suggest that T4 DNA in a nascent state is mandatory for the initial late gene transcription, or perhaps for late gene transcription throughout the lytic cycle. T4 DNA in a nascent state could be segregated from the bulk of the replicating DNA, used only as template for RNA synthesis, and prevented from being modified by methylation, glucosylation, or maturation processes. The fact that no, or very little, nonglucosylated T4 DNA is extractable from T4LB3-infected CR63 after arresting DNA synthesis does not rule out this possibility.
Functional Heterogeneity of Virions in Human Adenovirus Types 2 and 12
Authors: Rainbow Andrew J. , Mak Stanley
Published: January 2020

Purified preparations of adenovirus types 2 and 12 were used to infect KB cells at different input multiplicities. The resulting infected cultures were scored for inclusion body formation, production of infectious centers, and cloning efficiency. Both preparations were found to contain some defective particles capable of preventing a cell from cloning but unable to induce inclusion bodies or form plaques. The proportion of such defective particles in adenovirus 12 was about 10 times that in adenovirus 2. At high input multiplicities, the percentage of cells displaying an inclusion body was less than that predicted by the Poisson distribution and reached a maximum of 40 to 60% for adenovirus 2 and 12 to 15% for adenovirus 12. This reduction may be due to interference by large numbers of non-plaque-producing particles infecting each cell. The per cent of cells forming infectious centers was substantially greater for adenovirus 2 than for adenovirus 12 when compared at the same input plaque-forming units, reaching a maximum of 35 to 73% for adenovirus 2 and 5 to 10% for adenovirus 12. The low value for adenovirus 12 may be a result of the same interference phenomenon.
Host-Range Restrictions of Murine Leukemia Viruses in Mouse Embryo Cell Cultures
Authors: Hartley Janet W. , Rowe Wallace P. , Huebner Robert J.
Published: January 2020

Murine leukemia virus strains fall into three categories with respect to their ability to propagate in cells of National Institutes of Health (NIH) Swiss and BALB/c mouse embryos. Cultures of NIH cells are 100- to 1,000-fold more sensitive to “N-tropic” strains than BALB/c cell cultures, but are 30- to 100-fold less sensitive to “B-tropic” strains. Some virus strains (dually tropic or “NB-tropic”) propagate equally well in both cells. M-MSV pseudotypes show the host-range characteristics of the virus supplying the envelope, both in vitro and in vivo. The host-range characteristics appear to be genetically determined and could not be explained by host-induced modification or virus mixtures. There was no correlation between host range and Gross-AKR or FMR serotype.
Delay of Mengovirus-Induced Cytopathology in Mitotic L-Cells
Authors: Lake R. S. , Winkler D. C. , Ludwig E. H.
Published: January 2020

The time course and extent of mengovirus production were the same in metaphase-arrested and interphase L-cells, yet the expression of cytopathology was delayed several hours in metaphase cells.
Polyacetal Carboxylic Acids: a New Group of Antiviral Polyanions
Authors: Claes P. , Billiau A. , De Clercq E. , Desmyter J. , Schonne E. , Vanderhaeghe H. , De Somer P.
Published: January 2020

Chlorite-oxidized oxypolysaccharides are polyacetal carboxylic acids. They inhibited the cytopathic effect of vesicular stomatitis virus in mouse embryo cell cultures challenged at low input multiplicity. After intraperitoneal injection of these compounds in mice, interferon appeared in the circulation. The compounds also protected mice against lethal mengovirus infection and against the development of experimental pox lesions on the tail. Chlorite-oxidized oxyamylose was antiviral only when at least 64% of the glucopyranose units were oxidized, an observation which suggested a correlation between charge density and antiviral effect. The antiviral activity was also influenced by the molecular weight, as demonstrated by the fact that chlorite-oxidized dextrans which had a high intrinsic viscosity were more active than those with low intrinsic viscosity.
Density Differences Between Hybrid and Nonhybrid Particles in Two Adenovirus-Simian Virus 40 Hybrid Populations
Authors: Baum Stephen G. , Wiese William H. , Rowe Wallace P.
Published: January 2020

The adenovirus 7-simian virus 40 hybrid virus population E46
+
was subjected to fixed-angle equilibrium density gradient centrifugation in CsCl. A difference in buoyant density between the hybrid virion and its nonhybrid adenovirus 7 counterpart was noted, the hybrid virion possessing the lower buoyant density. This difference in buoyant density appeared to be accentuated in a population of adenovirus 2
+t7
, a derivative of E46
+
in which the adenovirus 7-simian virus 40 genome had been transferred to an adenovirus 2 capsid.
Biophysical Comparison of Two Canine Adenoviruses
Authors: Marusyk R. G. , Norrby E. , Lundqvist Ulla
Published: January 2020

The two canine adenoviruses, infectious canine hepatitis (ICH) virus and infectious canine laryngotracheitis (ICL) virus (also designated as Toronto A26/61 virus), were studied with respect to their morphology and the biological properties of their soluble components. The two viruses were found to be composed of soluble components similar to, and carrying biological activities corresponding to, those of the human adenoviruses after fractionation by rate zonal centrifugation and anion-exchange chromatography. The following soluble components were identified: hexons (carrying a common group-specific complement-fixing antigen), penton oligomers (complete soluble hemagglutinin), and penton and fiber monomers (incomplete soluble hemagglutinins). The latter were indicated by hemagglutination enhancement with selected antisera directed against human adenovirus soluble components. Elution characteristics of corresponding components of the two viruses in anion-exchange chromatography experiments were distinctly different. Electron microscopic examination of purified virions and soluble components revealed the fiber component of ICL virus to be 35 to 37 nm in length, and that of ICH virus to be 25 to 27 nm long.
Physical and Biological Properties of Dengue-2 Virus and Associated Antigens
Authors: Smith Thomas J. , Brandt Walter E. , Swanson John L. , McCown Jack M. , Buescher Edward L.
Published: January 2020

Dengue virus suspensions from mouse brain and cell culture were fractionated into three components by rate zonal centrifugation in sucrose gradients. Infectious virus sedimented in a single zone and possessed hemagglutinating (HA) and complement fixing (CF) activity. Electron micrographs showed the virion to be a spherical particle 48 to 50 nm in diameter with 7-nm spherical structures on its surface. Buoyant density in CsCl of virions from mouse brain was estimated at 1.22 g/cm
3
and from cell culture at 1.24 g/cm
3
. During centrifugation of virions in CsCl, an additional HA component appeared with a buoyant density of 1.18 g/cm
3
. It was shown in electron micrographs to consist of virion fragments. A noninfectious component with HA and CF activity sedimented in sucrose more slowly than intact virus, had a buoyant density of 1.23 g/cm
3
in CsCl, and appeared as “doughnut” forms measuring 13.8 to 14 nm in diameter. A third component, with CF activity and no HA activity, sedimented very little in sucrose gradients. Particles of the same size and shape as the spherical subunits on the surface of the virion were observed in electron micrographs.
Induction and Repression of
l
-Arabinose Isomerase in Bacteriophage-Infected
Salmonella typhimurium
Authors: Chakravorty Maharani
Published: January 2020

The induction of
l
-arabinose isomerase in
Salmonella typhimurium
(LT2) is repressed on infection with clear plaque forming mutants (C
1
and C
2
) of the temperate phage P22 (C
+
). However, after infection with C
+
leading to lysogeny, there is a temporary repression. During this period, messenger ribonucleic acid (RNA) for
l
-arabinose isomerase accumulates. DNA-RNA hybridization data suggest that there is transcription of host DNA during the period of repression. Interference at the level of translation might be responsible for the cessation of induced enzyme synthesis.
Morphological Changes in Productive and Abortive Infection by Feline Herpesvirus
Authors: Abraham Andrew , Tegtmeyer Peter
Published: January 2020

Feline herpesvirus produces characteristic morphological alterations in feline kidney cells. Nucleocapsid particles are formed in infected nuclei and are enveloped as they pass through the modified inner nuclear membrane. Aggregates of dense granular material and filamentous structures also regularly appear in infected nuclei. Infection of human embryonic lung cells by feline herpesvirus results in the appearance of intranuclear inclusion bodies, aggregates of dense granular material, and bundles of parallel filaments but no nucleocapsid particles.
Replication of Sendai Virus
Authors: Blair Carol D. , Robinson William S.
Published: January 2020

Chick embryo fibroblast cultures infected with Sendai virus were incubated with
3
H-uridine in the presence of actinomycin D beginning at 18 hr after infection. The 35 and 18
S
virus-specific ribonucleic acid (RNA) components were found in a ribonuclease-sensitive form in the cell and appeared to be associated with polyribosomes. Newly synthesized 57
S
viral RNA was rapidly coated with protein to form intracellular viral nucleocapsid, and no 57
S
RNA was found “free” (ribonucleasesensitive) in the 2,000 ×
g
supernatant fraction of disrupted cells. The nucleocapsid from detergent-disrupted Sendai virus and that from disrupted cells were indistinguishable in ultrastructure and buoyant density, and neither was found to be infectious or have hemagglutinating activity. Kinetic studies of nucleocapsid and virus formation indicated a relative block in conversion of viral nucleocapsid to complete enveloped virus in these cells, resulting in accumulation of large amounts of nucleocapsid in the cell cytoplasm.
Inhibition Phenomenon of Staphylococci by Bacteriophages
Authors: Chun Doki , Chung Jae Kyu
Published: January 2020

In the study of the relationship between bacteriophage and strains of staphylococci showing inhibition, slight differences were observed in the ability to adsorb phage between staphylococci of full phage sensitivity and those showing inhibition by phage. Only a few plaques were produced by inhibitory phages adsorbed on strains showing inhibition, whereas almost all of the phages adsorbed on corresponding phage-propagating strains produced plaques. Some strains showing inhibition were converted to full sensitivity to certain phages by heat shock or trypaflavine treatment. Treated strains adsorbed inhibitory phages to almost the same degree as nontreated strains, but most of the phages adsorbed on treated strains produced plaques. Killing was not always observed in cells adsorbing inhibitory phages. These results suggest that inhibition is not due to low adsorption rates, but rather to plaque formation by a small number of the sensitive fraction of the population and overgrowth by nonlysed cells.
Lymphotropic papovavirus transformation of hamster embryo cells
Authors: Takemoto K K , Kanda T
Published: January 2020

Hamster embryo cells were transformed by African green monkey lymphotropic papovavirus (LPV). The transformed cells contained intranuclear T-antigens demonstrable by fluorescent antibody staining with hamster anti-LPV serum. Analysis of uncloned and cloned lines of transformed cells for LPV sequences revealed that the viral DNA was present as free nonintegrated and integrated genomes; there were approximately 10 copies of free DNA and about one to two copies of integrated genomes per cell. The cells were highly tumorigenic when inoculated into hamsters and produced progressively growing tumors in 100% of newborn or 10-day-old hamsters that were inoculated with LPV-transformed cells. The serum from tumor-bearing hamsters reacted with LPV-transformed cells and also showed a weak reaction with simian virus 40-, BK virus-, and JC virus-transformed cells, thereby showing an antigenic relationship with the T-antigens of other primate polyomaviruses. The large T-antigen of LPV was found to be an 84,000-molecular-weight protein which was immunoprecipitated by hamster anti-LPV (antiviral) as well as by tumor serum.
Characterization of genetic changes occurring in attenuated poliovirus 2 during persistent infection in mouse central nervous systems
Authors: Rozhon E J , Wilson A K , Jubelt B
Published: January 2020

Genomic changes occurring in the attenuated W-2 strain of poliovirus 2 during persistent infection of the central nervous system of immunosuppressed mice were analyzed. The RNase T1 oligonucleotide fingerprints of 34 different viruses, isolated from the brains and spinal cords of paralyzed and nonparalyzed mice during a 105-day period, were used to quantitate and compare the mutations occurring in each isolate. Although mice were inoculated with plaque-purified virus, genetically distinct viruses were recovered from the central nervous system. The number of oligonucleotide changes occurring in isolates from paralyzed mice generally was greater than that observed in isolates from nonparalyzed mice. However, differences in the extent of mutation in isolates from the two groups of mice did not appear to be related to the level of virus replication. In paralyzed mice, the number of oligonucleotide changes on average was greater in viruses isolated during the first 60 days of the infection than in the last 45 days. The number of oligonucleotide changes was essentially constant throughout the infection, however, in viruses isolated from the brains of nonparalyzed mice. In addition, several specific oligonucleotide changes were found only in viruses isolated from paralyzed animals.
Free and integrated recombinant murine leukemia virus DNAs appear in preleukemic thymuses of AKR/J mice
Authors: Herr W , Gilbert W
Published: January 2020

We studied the appearance and structure of murine leukemia viral genomes in preleukemic AKR/J mice by Southern hybridization. Up to an average of one to two copies per thymocyte of unintegrated murine leukemia virus DNA appears in the thymuses of preleukemic mice beginning at 4 to 5 months of age and disappears in leukemic thymuses. The free viral genomes are absent in the spleens, livers, and brains of preleukemic mice. Using a series of ecotropic and nonecotropic murine leukemia virus hybridization probes, we showed that the unintegrated viral genomes are structurally analogous to those of recombinant mink cell focus-forming viruses that appear as proviruses in leukemic AKR thymocytes, suggesting that these free viral DNAs are the direct precursors to the leukemia-specific proviruses. The mosaic of ecotropic and nonecotropic sequences within these unintegrated viral DNAs varies from one preleukemic thymus to another but often appears structurally homogeneous within individual thymuses, indicating that often each thymus was being infected by a unique mink cell focus-forming virus. Analysis of high-molecular-weight DNA shows that recombinant proviruses reside in the chromosomal DNA of thymocytes within the preleukemic thymus, with the number rising to an average of several copies per thymocyte, but we do not detect any preferred integration sites. These results suggest that, in general, before the development of thymic leukemias in AKR mice there is a massive infection by a unique mink cell focus-forming virus which then integrates into many different sites of individual thymocytes, one of which grows out to become a tumor.
T4-induced antipolarity: temporal heterogeneity in response of early transcription units
Authors: Thermes C , Brody E
Published: January 2020

When T4 infects Escherichia coli in the absence of protein synthesis, rho-mediated termination takes place on early polycistronic transcription units. During the early period of development, the appearance of delayed early transcripts becomes insensitive to the inhibition of protein synthesis. In the absence of the T4 gene product mot, an inducer for the middle mode of transcription, only the early polycistronic messengers are synthesized. In mot- -infected cells, the synthesis of the distal transcripts still becomes completely insensitive to the polar effect of chloramphenicol. This happens because potential rho-sensitive termination sites are not used in these cells. In this respect, overcoming polarity induced by chloramphenicol can be called a process of antitermination. The mot-independent antitermination can be studied by addition of chloramphenicol during infections with mot- bacteriophage. The effect is stable; it allows a constant percentage of rho-sensitive termination sites in the cell to be traversed by RNA polymerase for at least 10 min at 42 degrees C. By examining six different transcription units on the T4 genome, we find that each transcription unit has a cis-acting component (or components) which determines when its rho-sensitive termination site stops functioning. In extreme cases, rho acts with 100% efficiency in some transcription units, whereas it is almost inactive in others.
Endogenous xenotropic murine leukemia virus-related sequences map to chromosomal regions encoding mouse lymphocyte antigens
Authors: Wejman J C , Taylor B A , Jenkins N A , Copeland N G
Published: January 2020

DNAs of all inbred mouse strains contain multiple copies (18 to 28 copies per haploid mouse genome) of endogenous xenotropic murine leukemia virus-related sequences detectable by Southern analysis with a xenotropic murine leukemia virus env gene-specific probe. After PvuII digestion, we identified a subset of xenotropic murine leukemia virus-related sequences that are well resolved by agarose gel electrophoresis and can be mapped to specific chromosomes by using recombinant inbred mouse strains. Interestingly, three of six xenotropic proviral loci that we mapped were integrated near genes encoding mouse lymphocyte antigens (Ly-m22, chromosome 1; Ly-m6, chromosome 2; and Ly-m10, chromosome 19) and a fourth xenotropic proviral locus mapped near a gene on chromosome 4 that has a major influence on xenotropic virus cell surface antigen levels. These studies indicate that xenotropic proviral loci are located on many different mouse chromosomes and may be useful markers for molecularly cloning and characterizing regions of the mouse genome important in lymphocyte development.
Effect of the beta-gamma phosphate bond of ATP on synthesis of leader RNA and mRNAs of vesicular stomatitis virus
Authors: Green T L , Emerson S U
Published: January 2020

RNA products synthesized in vitro by vesicular stomatitis virus with normal nucleotides or imido analogs were compared by polyacrylamide gel electrophoresis. When imido ATP, which has a nonhydrolyzable beta-gamma bond, was substituted for ATP, leader RNA and DI particle product were synthesized, but appreciable mRNA synthesis was not detected.
Prolonged inhibitory effect of 9-(1,3-dihydroxy-2-propoxymethyl)guanine against replication of Epstein-Barr virus
Authors: Lin J C , Smith M C , Pagano J S
Published: January 2020

The effects of 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG), a new antiviral drug, and acyclovir (ACV) [9-(2-hydroxyethoxymethyl)guanine] on the replication of Epstein-Barr virus (EBV) were compared. Both drugs inhibited EBV DNA replication in P3HR-1 cells and superinfected Raji cells, but neither inhibited replication of the plasmid form of the EBV genome in latently infected Raji cells. However, DHPG had a more prolonged inhibitory effect than ACV. Although the effect of the drugs is prompt, the kinetics of inhibition of EBV replication indicated that a drug exposure of 14 days was needed to reduce the EBV genome copy number to the residual plasmid level (30 copies per cell). The inhibitory effect of ACV was readily reversed within 11 days after removal of the drug, in contrast to the more prolonged effect exerted by DHPG, which persisted for more than 21 days. The 50% inhibitory doses for cell growth of ACV and DHPG were estimated to be 250 and 200 microM, respectively. The viral 50% and 90% effective doses of inhibition were, respectively, 0.3 and 9 microM for ACV and 0.05 and 3 microM for DHPG. The therapeutic indices (50% inhibitory dose/50% effective dose) for ACV and DHPG were 833 and 4,000, respectively. Synthesis of EBV-associated polypeptides was also affected. In superinfected Raji cells, ACV (100 microM) and DHPG (30 microM) inhibited synthesis of polypeptides with molecular weights of 145,000 and 140,000; in addition, synthesis of polypeptides with molecular weights of 110,000 and 85,000 was markedly reduced by DHPG but not by ACV. However, after drug removal, the inhibitory effect of ACV on polypeptide synthesis was abolished in contrast to the more persistent effect of DHPG.
Pathogenesis of in utero infections with abortogenic and non-abortogenic alphaviruses in mice
Authors: Milner A R , Marshall I D
Published: January 2020

The initial stages of infection of pregnant mice at gestation day 11 with either the T48 strain of Ross River virus or avirulent Semliki Forest virus are similar. With both infections, a hematogenous spread of virus to the placenta occurs. The viruses subsequently replicate to high titer in all placentas and are able to persist in the presence of specific maternal antiviral antibodies. There is a delay of at least 1 to 2 days between the initial detection of virus in the placenta and the onset of fetal infection. With Semliki Forest virus, abortion occurred in all mothers and appeared to be preceded by infection of all fetuses. However, when Semliki Forest virus was given at other stages of pregnancy, abortion was less common, and in all non-aborted pregnancies at least one uninfected fetus was observed. This situation was similar to that with Ross River virus, in which abortion was not observed and fetal infection and death were only seen in a proportion of fetuses. Within each pregnancy, the outcome of the two in utero infections appeared to result from similar mechanisms, with the fate of an individual fetus depending upon the timing of the passive transfer of anti-viral immunoglobulin G from the mother relative to the timing of fetal infection by virus from the placenta. Although the passive maternal immunoglobulin G protected susceptible fetuses against infection, antibody did not mediate in utero recovery of infected fetuses or clear placental infection.
Characterization of the DNA-protein complex at the termini of the bacteriophage PRD1 genome
Authors: Bamford D H , Mindich L
Published: January 2020

DNA of bacteriophage PRD1 has protein P8 at its termini. Extracts of infected cells are able to derivatize P8 in vitro with labeled dGTP. Two early proteins, P1 and P8, products of genes I and VIII, respectively, are the only phage proteins necessary for the formation of the protein P8-dGMP complex. This was shown by complementation of extracts from cells infected with mutants and by use of extracts from cells carrying cloned genes I and VIII. With Escherichia coli mutants that are temperature sensitive for DNA synthesis, it was possible to show that the formation of the protein P8-dGMP complex was dependent upon the host replication apparatus. The analysis of the purified protein P8-dGMP complex by hydrolysis and enzymatic digestion showed that there is a covalent phosphodiester bond between tyrosine and 5'-dGMP.
Expression of hepatitis B antigens with a simian virus 40 vector
Authors: Will H , Cattaneo R , Pfaff E , Kuhn C , Roggendorf M , Schaller H
Published: January 2020

Recombinant DNA molecules consisting of the simian virus 40 (SV40) early region and different subgenomic hepatitis B virus DNA fragments were constructed in vitro and packaged in vivo into SV40 capsids by using a complementing SV40 helper virus. Upon infection with these virus stocks the three known hepatitis B-specific antigens were expressed under SV40 control. The surface antigen was released into the medium, and the core antigen and its derivative hepatitis B e antigen were only detected intracellularly. Size analysis of the core gene product(s) by immunoblotting revealed the presence of a single protein species identical with the 21-kilodalton core antigen isolated from human liver. The hepatitis B core antigen expressing construct did not contain a putative precore sequence, indicating that such a sequence is not needed for hepatitis B core antigen synthesis in animal cells. S1 analysis demonstrated the use of SV40 signals for initiation and polyadenylation of the core gene transcripts. In addition, a processing-polyadenylation signal was identified within the core gene.
Molecular cloning and long terminal repeat sequences of intracisternal A-particle genes in Mus caroli
Authors: Ono M , Kitasato H , Ohishi H , Motobayashi-Nakajima Y
Published: January 2020

We isolated DNA clones of intracisternal A-particle (IAP) genes from the genome of an Asian wild mouse, Mus caroli. A typical M. caroli IAP gene was 6.5 kilobase pairs in length and had long terminal repeat (LTR) sequences at both ends. The size of the LTR was 345 base pairs in clone L20, and two LTRs at both ends of this clone were linked to directly repeating cellular sequences of 6 base pairs. Each LTR possessed most of the structural features commonly associated with the retrovirus LTR. The restriction map of the M. caroli IAP gene resembled that of Mus musculus, although the M. caroli IAP gene was 0.4 kilobase pairs shorter than the M. musculus IAP gene in two regions. Sequence homology between the M. caroli and M. musculus IAP LTRs was calculated as about 80%, whereas the LTR sequence of the Syrian hamster IAP gene was about 60% homologous to the M. caroli LTR. The reiteration frequency of the M. caroli IAP genes was estimated as 200 to 400 copies per haploid genome, which is at least 10 times the reported value. These results suggest that the IAP genes observed in the genus Mus are present in multiple copies with structures closely resembling the integrated retrovirus gene.
Immunobiochemical characterization with monoclonal antibodies of Epstein-Barr virus-associated early antigens in chemically induced cells
Authors: Epstein A L
Published: January 2020

Five monoclonal antibodies which are reactive to early antigens of Epstein-Barr virus have been produced by using somatic cell hybridization techniques. The specificity of the monoclonal antibodies to early antigens was demonstrated by indirect immunofluorescence, which showed that the antigens were localized to the nucleus of early antigen-induced Raji cells. Additional indirect immunofluorescence studies showed that like patient antisera to diffuse-staining early antigen, the monoclonal antibodies gave positive staining reactions after methanol fixation. One of the antibodies, 1150-4, was positive by the anti-complement immunofluorescence technique but differed with Epstein-Barr virus-associated nuclear antigen-positive patient sera in that it only stained induced cells. Different fixation methods were found to alter dramatically the appearance of the nuclear staining reactions produced by the monoclonal antibodies. Immunoprecipitation and immunoblot experiments revealed that monoclonal antibodies 1108-1 and 1129-1 recognized two polypeptides of 55,000 and 50,000 daltons (p55;50), 1173-6 and 1180-2 recognized just p50, and 1150-4 identified a 65,000-dalton nuclear protein. Immunobiochemical characterization of these viral antigens showed that p55 is a phosphoprotein, and p55;50 has strong DNA-binding activity preferentially to single-stranded DNA. Elucidation of the role of these nuclear proteins in Epstein-Barr virus infection and the events associated with Epstein-Barr virus-directed lymphocyte transformation may provide significant information on the pathogenicity of this important human virus.