bluuebunny/crossref_metadata_2025 huggingface dataset:
Human cytomegalovirus: glycoproteins associated with virions and dense bodies
Authors: Stinski M F
Published: January 2020

The glycoproteins associated with the membranes of cytomegalovirions and dense bodies were characterized by their relative mobility, percentage of glucosamine incorporation, and molecular weight. Eight glycopolypeptides were repeatedly detectable. Three glycopolypeptides of higher molecular weight with low levels of glucosamine incorporation were occasionally detectable. These latter glycopolypeptides may be precursors or aggregates of the glycopolypeptides with lower molecular weights. The glycoproteins associated with the membranes were on the surface, as determined by iodination with 125I of virions and dense bodies partially purified in gradients of D-sorbitol. Velocity centrifugation in linear gradients of D-sorbitol was used to obtain concentrated and partially purified preparations of infectious cytomegalovirus. Viral infectivity and the membranes of cytomegalovirions and dense bodies were stable in gradients of sorbitol, but cellular contaminants were not completely removed. Additional centrifugation in CsCl separated both cellular contaminants and viral nucleocapsids from virions and dense bodies. Many dense bodies, which are considered to be aberrant forms of cytomegalovirus, had the same size, sedimentation properties, and density as virions. Consequently, they were not separable from virions by various centrifugation techniques. Electron microscopy demonstrated that purified virions and dense bodies were qualitatively free of extraneous material and that each dense body was bounded by a membrane, as evidenced by its double-tract appearance. Antisera to a preparation of purified virions and dense bodies, or to their glycoproteins, contained antibodies that neutralized viral infectivity and reacted with antigens in cells infected with cytomegalovirus. However, these same antisera did not contain antibodies that reacted with uninfected cells. The glycoproteins associated with the membranes of cytomegalovirions and dense bodies are considered to be specified by the cytomegalovirus genome.
Polyadenylic acid in the genomic RNA of mengovirus
Authors: Marshall M V , Arlinghaus R B
Published: January 2020

The polyadenylic acid contained in 35S mengovirus RNA produced in infected BHK-21 cells contained approximately 94% AMP and was estimated to contain an average of 50 to 55 nucleotides. The polyadenylic acid is placed at the 3'-end of the genomic RNA based on the presence of significant levels of [3H]adenosine in complete alkali or RNase T2 digests of polyadenylic acid from [3H]adenosine-labeled 35S viral RNA.
Induction of murine p30 by superinfecting herpesviruses
Authors: Reed C L , Rapp F
Published: January 2020

The interaction of endogenous type C viruses with superinfecting herpes simplex virus type 2 (HSV-2) was investigated in two murine cell lines. Replication of HSV-2 was suboptimal in random-bred Swiss/3T3A cells and, in initial experiments, infection with a low virus-to-cell ratio resulted in carrier cultures with enhanced murine leukemia virus (MuLV) p30 expression. Immunofluorescence tests with Swiss/3T3A cells productively infected with HSV-2 also showed HSV-associated cytoplasmic antigens and enhanced MuLV p30 expression when compared with uninfected controls. Inactivation of HSV-2 with UV light did not abolish this reaction, although the number of cells expressing p30 was reduced. HSV-2 replicated more efficiently in a line of NIH Swiss cells (N c1 A c1 10). These cells are not readily inducible for type C expression by conventional methods; however, untreated and UV-inactivated HSV-2 induced both HSV-2-associated antigens and MuLV p30 in these cells. Although the Birch strain of human cytomegalovirus induced MuLV p30, neither mouse cytomegalovirus nor vesicular stomatitis virus induced MuLV p30 in either cell line.
Genome localization of adeno-associated virus RNA
Authors: Carter B J , Fife K H , de la Maza L M , Berns K I
Published: January 2020

In previous work, linear duplex molecules of adeno-associated virus, type 2 (AAV2), DNA were cleaved with the restriction endonucleases R-EcoRI, R-HindII, and R-HindIII. The physical order of the specific fragments obtained was deduced and oriented with respect to the DNA strand polarity and the direction of transcription. Stable AAV RNA is transcribed only from 70% of the minus DNA strand. We report here RNA-DNA hybridization experiments using these restriction fragments to obtain a more accurate map of the portion of the AAV genome represented in stable RNA. The data obtained with several sets of restriction fragments annealed to either whole-cell RNA or poly(A)-containing RNA were internally consistent. The AAV RNA annealed with a continuous region of the AAV DNA, beginning at 0.18 map units (18%) from the left end of the molecule and ending at 0.88 map units. In addition, the restriction endonuclease BamHI was found to make one specific cleavage in AAV2 DNA at 0.22 map units, which is 0.04 map units (i.e., 160 nucleotides) to the right (""down stream'') of the point corresponding to the 5' end of the viral mRNA.
Characteristics of Sindbis virus temperature-sensitive mutants in cultured BHK-21 and Aedes albopictus (Mosquito) cells
Authors: Renz D , Brown D T
Published: January 2020

A number of the temperature-sensitive mutants of Sindbis virus originally isolated and characterized by Burge and Pfefferkorn (1966, 1968) were reexamined for their abilities to grow and complement one another in cultured BHK-21 and Aedes albopictus (mosquito) cells. The response of the mutants to conditions of high and low temperature was similar in cultured cells of both the vertebrate and invertebrate hosts. Complementation experiments in BHK-21 cells produced growth patterns similar to those described by Burge and Pfefferkorn for chicken embryo fibroblast cells (1966) and placed the mutants into six nonoverlapping complementation groups. When examined in the cultured mosquito cells, only three of the nine mutants used in this study demonstrated complementation under a variety of experimental conditions. Homologous interference experiments demonstrated that the unusual patterns of complementation obtained in the A. albopictus cells did not result from an inefficient infection of the invertebrate cells by the mutants.
Characterization of an RNA-a protein complex isolated from the RNA bacteriophage M12
Authors: Leipold B , Hofschneider P H
Published: January 2020

The properties of an RNA-A protein complex isolated from the RNA bacteriophage M12 are described. The molar ratio of RNA to A protein in the complex is estimated to be 1:1. In sucrose gradients, the complex sediments like free RNA molecules. In contrast to RNA alone, which can only infect spheroplasts, the RNA-A protein complex infects intact Escherichia coli cells and produces infectious progeny particles like the original phage. Evidence is presented that the infection of the host cells by the complex takes place via F pili. All of the infectivity disappears if the ionic bonds of RNA to A protein in the complex are dissociated in 0.5 M sodium chloride buffer at 37 degrees C. Furthermore, the kinetics of complex dissociation and loss of infectivity are the same, implying that the binding of A protein to the RNA is a prerequisite for infectivity on intact host cells.
Glycosylation sites of vesicular stomatitis virus glycoprotein
Authors: Robertson J S , Etchison J R , Summers D F
Published: January 2020

Detailed analysis on DEAE-Sephadex of the tryptic digestion products of the glycoprotein from vesicular stomatitis virus grown in HeLa suspension cultures revealed the presence of two major and several minor sugar-labeled species. The minor tryptic glycopeptides were converted to one of the two major glycopeptide species by treatment with neuraminidase. Thus, vesicular stomatitis virus glycoprotein contains only two oligosaccharide side chains that are heterogeneous in their sialic acid content.
Physical and chemical characterization of an avian reovirus
Authors: Spandidos D A , Graham A F
Published: January 2020

The avian viral agent S1133 has previously been classified serologically as a member of the avian reovirus group. This viral agent grows in chicken embryo fibroblast cells, bands at a density of 1.37 g/ml in CsCl equilibrium density gradients, has a particle diameter of 75 nm, and has a morphology similar to that of human reovirus type 3. Its nucleic acid is comprised of double-stranded RNA and adenosine-rich oligonucleotides. The dsRNA is distributed among 10 segments with molecular weights of 2.7 x 10(6), 2.6 x 10(6), 1.7 x 10(6), 1.5 x 10(6), 1.3 x 10(6), 1.2 x 10(6), 0.80 x 10(6), 0.74 x 10(6), and 0.68 x 10(6) for the largest (L1) to the smallest (S4) segment, respectively, as determined by polyacrylamide gel electrophoresis. These 10 segments migrate differently on polyacrylamide gels compared to those of human reovirus type 3. The capsid proteins of avian reovirus consist of eight species of polypeptides as determined by polyacrylamide gel electrophoresis. These are lambda1, lambda2, lambda3, mu1, mu2, sigma1, sigma2, and sigma3 with molecular weights of 140, 125, 115, 85, 72, 40, 36, and 32 x 10(3), respectively. Only polypeptide sigma2, which resides in the inner capsid or core, comigrated with the sigma2 polypeptide of type 3 reovirus. Antiserum against type 3 reovirus did not neutralize avian reovirus. Avian reovirus core particles were found to possess a transcriptase and a methylase activity.
Inhibition of Host-Cell Protein and Ribonucleic Acid Synthesis by Newcastle Disease Virus
Authors: Wilson D. E.
Published: January 2020

The mechanisms of Newcastle disease virus-(NDV) induced inhibition of cell protein and ribonucleic acid (RNA) synthesis were investigated. It was observed that the ability of NDV to inhibit cell RNA synthesis is dependent on the virus strain. The inhibitors, azauridine and cycloheximide, were added to cell cultures at different times after infection to study the roles of protein and RNA synthesis in the viral inhibition process. Viral inhibition of cell RNA synthesis and viral inhibition of cell protein synthesis become resistant to cycloheximide at a different time after infection than that in which they become resistant to azauridine. The results indicate that the inhibition of cell RNA synthesis by the Texas strain involves the synthesis of inhibitory proteins which are coded by the viral genome. The Texas and Beaudette strains of NDV appear to employ different mechanisms for the inhibition of host-cell protein synthesis. Viral inhibition of cell protein synthesis does not appear to cause, or be the result of, viral inhibition of cell RNA synthesis.
Ultrastructure of Bacterial Cells Infected with Bacteriophage PM2, a Lipid-containing Bacterial Virus
Authors: Cota-Robles Eugene , Espejo Romilio Torres , Haywood Patricia Williams
Published: January 2020

The cytological pattern of infection of a host pseudomonad with PM2, a lipid-containing bacterial virus, was investigated by electron microscopy. Normal and infected cells frequently contain a myelin figure, which is found in the nucleoid region or at the periphery of the cell. The most striking finding in this investigation was that completed virions are found in the cell adjacent to or in association with the cytoplasmic membrane. This localization is precise; virions are not found elsewhere in infected cells. The completed virions occasionally appear to be attached to the cytoplasmic membrane. The virus contains a darkly staining core surrounded by a tripartite envelope of a thickness of approximately 70 A, which is identical to the thickness of the cytoplasmic membrane. Lysing cells appear to undergo extensive damage of the cytoplasmic membrane prior to rupture of the L layer of the cell wall.
Inhibition of Arbovirus Protein Synthesis by Interferon
Authors: Friedman Robert M.
Published: January 2020

Infection of cells treated with guanidine and actinomycin D and then washed to remove the guanidine inhibition of virus growth had no effect on antiviral activity already established by interferon. Protein synthesis in interferon-treated cells infected under these conditions was decreased as compared to control cells similarly treated but not exposed to interferon. In these control cells, analysis by polyacrylamide gel electrophoresis indicated that six proteins were produced during the first hour after guanidine reversal. Five of these proteins have been previously identified as probably being viral in origin. In interferon-treated cells, only a single major protein was produced. Ribonucleic acid (RNA) synthesis by Semliki Forest virus during the first hour after guanidine reversal was markedly depressed by incubation at 42 C, but no inhibition of total virus protein synthesis was seen; this finding suggested that much of the virus protein produced in the first hour after guanidine reversal was carried out by input virus RNA. Interferon was fully active in cells incubated at 42 C. The results suggested that interferon inhibits the production of Semliki Forest virus proteins ordinarily produced under the direction of the virus genome.
Cleavage by Formamide of Intercapsomer Bonds in Adenovirus Types 4 and 7 Virions and Hemagglutinins
Authors: Neurath A. Robert , Rubin Benjamin A. , Stasny John T.
Published: January 2020

When crude preparations of adenovirus types 4 and 7 were treated with solutions of formamide, a complete or nearly complete inactivation of infectivity could be achieved without affecting the complement fixation titers of the same preparations. Studies on purified virions showed that the intersubunit bonds within the viral capsid were broken and single capsomers were released as the result of treatment by formamide. When subjected to the same treatment, hemagglutinins were sequentially converted into incomplete hemagglutinins (pentons), which were in turn converted into penton bases and fibers.
Biophysical, Biological, and Cytochemical Features of a Virus Associated with Transplantable Hamster Tumors
Authors: Stenback W. A. , Van Hoosier G. L. , Trentin J. J.
Published: January 2020

A virus resembling type C murine leukemia viruses, which is associated with transplantable hamster tumors, was partially characterized with respect to certain biological, biophysical, and cytochemical features. As determined by electron microscopy, high concentrations of the virus appeared in the blood plasmas of tumor-bearing hamsters. Hamsters inoculated with virus concentrates did not show gross evidence of disease, and preliminary attempts to infect various hamster cells in tissue culture were unsuccessful. A line of cells from a virus-containing tumor which had been established in tissue culture released large numbers of the virus into the supernatant fluids by budding. The buoyant density peak of virus concentrates in potassium tartrate density gradients was 1.13 g/cm
3
by ultraviolet absorption and electron microscopic analysis. Acridine orange staining and nuclease digestion methods have indicated that the virus is probably a ribonucleic acid virus.
Comparative Studies on the Soluble Components of Adenovirus Types 9 and 15 and the Intermediate Strain 9-15
Authors: Norby Erling
Published: January 2020

Five different soluble components of adenovirus types 9, 9-15, and 15 have been identified. These are: (i) a slowly sedimenting, trypsin-resistant, incomplete hemagglutinin (HA). (This component was demonstrable by hemagglutination-enhancement (HE) tests in the presence of heterotypic antisera against members of Rosen's subgroups II and III, but
not
of subgroup I); (ii) a slowly sedimenting, trypsin-resistant, complete HA, causing only a partial agglutination of cells; (iii) a rapidly sedimenting, incomplete HA, demonstrable by HE tests in the presence of heterotypic antisera against members of all Rosen's subgroups. (Trypsin treatment of this component caused a conversion into slowly sedimenting incomplete HA); (iv) a group-specific complement-fixing (CF) antigen devoid of HA activity; and (v) a rapidly sedimenting, trypsin-sensitive, complete HA, which in the electron microscope was found to represent a dodecahedral aggregate of 12 pentons (a dodecon). On the basis of their biological and physicochemical characteristics, the first four components were interpreted to represent (i) fibers, (ii) a polymer of a few, probably two, fibers, (iii) pentons, and (iv) hexons, respectively. The length of fibers extending from dodecons and virions was estimated to be 11 to 14 nm. A similar value was suggested from exclusion chromatography experiments. Adenovirus types 9 and 15 fibers were recovered in a position intermediate to that of fibers of types 3 and 4, the lengths of which are 10 and 17 nm, respectively. The sequence of elution of different components of types 9 and 9-15 from an anion exchanger was fibers, fiber-aggregate, pentons, hexons, and dodecons. Type 15 components appeared in the same order except for the fact that dodecons eluted before hexons. The molarities of NaCl required to elute the different types 9 and 9-15 components, excluding hexons, were identical. They were distinctly different from those of the corresponding type 15 components. However, hexons of all three serotypes eluted in proximity to each other and there was a slight tendency for type 9-15 hexons to take a position intermediate to those of types 9 and 15.
Replication of Coliphage M-13 II. Intracellular Deoxyribonucleic Acid Forms Associated with M-13 Infection of Mitomycin C-treated Cells
Authors: Brown Lyle R. , Dowell C. E.
Published: January 2020

Intracellular deoxyribonucleic acid (DNA) forms associated with bacteriophage M-13 infection have been isolated and characterized.
Escherichia coli
HF4704 (F
+
,
hcr
−
,
thy
−
) cells were treated with mitomycin C to inhibit host-cell DNA synthesis and were then infected with phage M-13. This treatment permitted radioactive labeling of phage-specific DNA forms with
3
H-thymine. These labeled DNA components were characterized by sucrose density sedimentation and equilibrium density gradient centrifugation in neutral and ethidium bromide CsCl gradient. Two double-stranded circular forms were found with properties analogous to the replicative form I and replicative form II of φX174. A third component, identified as single-stranded DNA, was isolated in some samples removed 45 min after phage synthesis was initiated.
Structural Proteins of Reoviruses
Authors: Loh P. C. , Shatkin A. J.
Published: January 2020

Polyacrylamide gel electrophoresis of the solubilized proteins from the three serotypes of reovirus revealed that each contained three major and four minor components. Subviral particles were prepared by brief treatment of complete virions with urea. Electron microscopy, density-gradient centrifugation, and chemical analyses of these particles indicated that their outer capsid structure had been selectively removed. They contained only two proteins, but their ribonucleic acid composition was similar to that of complete virions. The subviral particles were not infectious.
Labile Coat: Reason for Noninfectious Cell-free Varicella-Zoster Virus in Culture
Authors: Cook Margery L. , Stevens Jack G.
Published: January 2020

Experiments designed to determine why cell-free varicella-zoster virus replicated in cell culture is noninfectious were performed. Electron micrographs in which varicella-zoster virus (a herpesvirus) was compared to herpes simplex virus in primary human amnion cell cultures showed that the viruses were morphologically indistinguishable inside the nucleus. However, extranuclear varicella-zoster viruses were distinguished from herpes simplex virus by the presence of pleomorphism, incomplete coats, and a resultant loss of central dense cores. This result indicates that varicella-zoster virus possesses a labile coat which is degraded outside the nucleus. It is suggested that the labile coat is a principal reason for the lack of cell-free infectious virus in this system.
Production of Virus by Mammalian Cells Transformed by Rous Sarcoma and Murine Sarcoma Viruses
Authors: Valentine Artrice F. , Bader John P.
Published: January 2020

Cultured cells of mammalian tumors induced by ribonucleic acid (RNA)-containing oncogenic viruses were examined for production of virus. The cell lines were established from tumors induced in rats and hamsters with either Rous sarcoma virus (Schmidt-Ruppin or Bryan strains) or murine sarcoma virus (Moloney strain). When culture fluids from each of the cell lines were examined for transforming activity or production of progeny virus, none of the cell lines was found to be infectious. However, electron microscopic examination of the various cell lines revealed the presence of particles in the rat cells transformed by either Rous sarcoma virus or murine sarcoma virus. These particles, morphologically similar to those associated with murine leukemias, were found both in the extracellular fluid concentrates and in whole-cell preparations. In the latter, they were seen budding from the cell membranes or lying in the intercellular spaces. No viruslike particles were seen in preparations from hamster tumors. Exposure of the rat cells to
3
H-uridine resulted in the appearance of labeled particles with densities in sucrose gradients typical of virus (1.16 g/ml.). RNA of high molecular weight was extracted from these particles, and double-labeling experiments showed that this RNA sedimented at the same rate as RNA extracted from Rous sarcoma virus. None of the hamster cell lines gave radioactive peaks in the virus density range, and no extractable high molecular weight RNA was found. These studies suggest that the murine sarcoma virus produces an infection analogous to certain “defective” strains of Rous sarcoma virus, in that particles produced by infected cells have a low efficiency of infection. The control of the host cell over the production and properties of the RNA-containing tumorigenic viruses is discussed.
Antiviral Activity of Antiserum Specific for an Influenza Virus Neuraminidase
Authors: Kilbourne E. D. , Laver W. G. , Schulman J. L. , Webster R. G.
Published: January 2020

Antiserum specific for influenza A
2
neuraminidase was produced by immunization of rabbits with the purified enzyme which had been isolated by electrophoresis from the proteins of a detergent-disrupted A
0
A
2
influenza virus recombinant [X-7 (F1)]. This recombinant contained hemagglutinin of the A
0
subtype and A
2
neuraminidase. Antiserum to the isolated A
2
neuraminidase did not react in any of four serological tests with A
0
or A
2
subtype viruses that lacked the A
2
enzyme. In contrast, the antiserum inhibited the neuraminidase activity only of wild-type and recombinant viruses containing the A
2
enzyme, regardless of the nature of their hemagglutinin proteins. The antiserum caused hemagglutination-inhibition of some, but not all, viruses bearing the A
2
enzyme, and it reduced the plaque size or plaque number of all viruses tested that contained A
2
neuraminidase. In the chick embryo and in cell culture, low dilutions of antiserum reduced the yield of virus. True neutralization of virus in the chick embryo did not occur. We conclude that an antiserum specific for A
2
neuraminidase influenced the yield and release of virus from influenza virus-infected cells.
Structure and Leukemogenic Activity of a Murine Leukemia Virus
Authors: de Tkaczevski L. , de Harven E. , Friend C.
Published: January 2020

Purified Friend viruses obtained from chronically infected tissue cultures were studied under the electron microscope in an effort to correlate the fine structure of the particles to their leukemogenic activity under varied experimental conditions, i.e., temperature treatments and exposure to Tween 80, amyl acetate, or ether. It was observed that an intact viral envelope was a prerequisite to leukemogenic activity as tested by intraperitoneal inoculation of newborn mice. It was also noted that the percentage of C particles was not increased after heating for 1 hr at 45 C (treatment which, however, completely inactivated the viruses). Digestion with ribonuclease indicated the presence of ribonucleic acid within the nucleoids of “enveloped A particles,” which shows that these are not immature particles. The significance of the simultaneous presence of “enveloped A” and C particles is discussed.
Mechanism of Photoreactivation of Pseudorabies Virus
Authors: Pfefferkorn E. R. , Coady Helen M.
Published: January 2020

Primary chick embryo cultures were able to photoreactivate ultraviolet-treated pseudorabies virus. Upon exposure to fluorescent light, infected or uninfected chick cells eliminated thymine dimers induced in their deoxyribonucleic acid by ultraviolet irradiation. In contrast, rabbit kidney cells did not photoreactivate the virus or eliminate thymine dimers. Thus, the capacity for photoreactivation appeared to be determined by the ability of the cell to eliminate thymine dimers.
Hybrid of Monkey and Human Adenoviruses
Authors: Altstein A. D. , Dodonova N. N. , Vassilyeva N. N. , Tsetlin E. M.
Published: January 2020

Following joint replication of monkey SA7 adenovirus (C8 strain) and human adenovirus type 2 in green monkey kidney tissue culture, a virus possessing the properties of a hybrid was obtained. It was designated Ad2C8. Ad2C8 preparations contained two types of viral particles: human adenovirus type 2, and hybrid particles. The hybrid virions multiplied in green monkey kidney cells in the presence of human adenovirus types 1, 2, and 3, but not 3 and 7, and acquired the capsid of the helper adenovirus. The hybrid can serve as a helper for human adenoviruses. It can apparently induce T antigen of the C8 virus but, in contrast to the latter, does not induce tumors in hamsters.
Electron Microscopy of Herpes Simplex Virus
Authors: Morgan Councilman , Rose Harry M. , Mednis Baiba
Published: January 2020

Although capsids of herpes simplex virus were encountered within phagocytic vesicles, they were more commonly observed free within the cytoplasm. Stages in the release of virus from vesicles were not seen. There appeared to be five distinct steps in the process whereby the virus initiates infection: attachment, digestion of the viral envelope, digestion of the cell wall, passage of the capsid directly into the cytoplasm, and digestion of the capsid with release of the core. Antibody probably interferes with the first two stages.
Mechanism of Resistance to Enteroviruses of Some Primate Cells in Tissue Culture
Authors: Soloviev V. D. , Krispin T. I. , Zaslavsky V. G. , Agol V. I.
Published: January 2020

Two cell lines, M10-45-2 and L-41, were studied, each of which possessed specific resistance either to poliovirus or to coxsackievirus. Infection of M10-45-2 cells with poliovirus ribonucleic acid (RNA) and L-41 cells with infectious coxsackievirus RNA was accompanied by production of complete viruses in each of the resistant cell lines. During incubation of the cells with the virus to which they were resistant, the amount of infectious virus did not decrease. Treatment with glycine-HCl buffer solution (
p
H 2.5) of resistant M10-45-2 cells after incubation with poliovirus at 0 C did not result in recovery of infectious virus, although such release did take place after treatment of sensitive M10 cells. Infection of resistant cells with virus containing poliovirus RNA and coxsackievirus proteins resulted in production of poliovirus in M10-45-2 cells but not in L-41 cells. The resistant cells are apparently unable to adsorb the virus to which they are resistant.
In Vitro Chick Embryo Cell Response to Strain MC29 Avian Leukosis Virus
Authors: Bolognesi D. P. , Langlois A. J. , Sverak L. , Bonar R. A. , Beard J. W.
Published: January 2020

Strain MC29 avian leukosis (myelocytomatosis) virus induced infection, elaboration of virus, and morphological alteration in chick embryo cells in vitro. Virus liberation began within 18 hr, morphological change was detectable at about 40 hr, and the cultures could be completely altered within 80 hr after infection. Altered cells were about half the volume and grew at approximately twice the rate of uninfected elements. The output of virus estimated by electron microscopy was about 140 particles per cell per hr. Deoxyribonucleic acid remained constant, but ribonucleic acid increased in both infected and control cells in adjustment to culture environment. The rates of uptake and incorporation of
3
H-uridine and the incorporation of
3
H-thymidine increased in the infected cells with onset of morphological change but were unaffected by processes of infection and virus elaboration per se. Incorporation of a
14
C-amino acid mixture was slightly greater in the infected than in control cells. The speed of continuity of infection and massive morphological alteration constitute a unique response to avian tumor viruses, and the system gives promise of singular value for detailed studies of the processes of infection and morphological change.
Relationship Between Competence for Transfection and for Transformation
Authors: Riva Silvano , Polsinelli Mario
Published: January 2020

Deoxyribonucleic acid (DNA) extracted from phage SPP1 is highly infectious on
Bacillus subtilis
competent cells; the efficiency of infection is 5 × 10
3
to 6 × 10
3
phage equivalents per plaque-forming unit. This DNA was used to study the relationship between competence for transfection and for transformation. The experiments were concerned with the frequency of infection and transformation in mutants exhibiting different levels of competence, the effect of periodate on competence for infection and for transformation, the competition between phage and bacterial DNA, the transformation of cells preinfected with phage DNA, and the infection of cells pretreated with bacterial DNA. The data show that
B. subtilis
cells competent for transformation are also competent for transfection and vice versa; transfection with phage DNA represents, therefore, a simple way to measure the total number of competent cells in a culture. The fraction of competent cells, determined by SPP1 DNA infection, varied from 10
−2
to 7 × 10
−2
.
Factors Affecting Infection of Protoplasts with Deoxyribonucleic Acid of Actinophage PK-66
Authors: Okanishi Masanori , Hamana Koei , Umezawa Hamao
Published: January 2020

To establish a method for transmission of genetic materials in the genus
Streptomyces
, the conditions of infection of protoplasts of
S. kanamyceticus
by actinophage PK-66 deoxyribonucleic acid (DNA) were studied. The protoplasts of
Streptomyces
were prepared by treatments with lysozyme and trypsin. The infectivity of the phage DNA was enhanced by the presence of NaCl in the medium. The optimal concentration of the protoplasts for infection with DNA was 7 × 10
7
to 4 × 10
8
/ml. A proportional relationship was found between the infectivity and the DNA concentration within a certain range. The maximal production of mature phage was achieved after 19 hr of incubation. The number of phage propagated in the infection mixture reached 10
4
plaque-forming units per ml under the appropriate conditions. The phage DNA infected not only protoplasts prepared from
S. kanamyceticus
but also those prepared from
S. violaceoniger
and
S. acidomyceticus
, which were resistant to intact phage PK-66.
Mechanisms of Enhancement of SP82 Transfection
Authors: Epstein H. T. , Mahler Inga
Published: January 2020

SP82 transfection in
Bacillus subtilis
could be markedly increased by exposing the competent cells to ultraviolet (UV)-irradiated homologous or heterologous cellular deoxyribonucleic acid (DNA). This enhancement was similar in time and level of peak effect to enhancements effected by preinfection with helper phages or by UV irradiation of competent cells. The effectiveness of various DNA preparations in increasing transfection paralleled the adenine plus thymidine content of the preparations and was maximal at UV doses approaching those which were maximal for pyrimidine dimerization. The most probable interpretation is that irradiated DNA binds cellular nucleases which would otherwise inactivate the incoming transfecting DNA.
Kinetics of Viral Deoxyribonucleic Acid, Protein, and Infectious Particle Production and Alterations in Host Macromolecular Syntheses in Equine Abortion (Herpes) Virus-infected Cells
Authors: O'Callaghan Dennis J. , Hyde James M. , Gentry Glenn A. , Randall Charles C.
Published: January 2020

Infection of exponential-phase suspension cultures of mouse fibroblast cells (L-M) with equine abortion virus (EAV) resulted in inhibition of cell growth and marked alterations in host metabolic processes. The synthesis of deoxyribonucleic acid (DNA) and ribonucleic acid was inhibited within 4 hr after infection and was suppressed by more than 90% by the time of maximal virus replication (14 to 18 hr). The overall rate of protein synthesis, however, was similar in uninfected and virus-producing cells as determined by measurements of net protein and isotope incorporation. The time course of viral DNA and protein synthesis and assembly into mature virus was determined with the inhibitors 5-fluorodeoxyuridine (FUdR) and cycloheximide, respectively. Thus, viral DNA synthesis was essentially completed at 14 hr, and viral protein and infectious virus synthesis was completed at 18 hr. Although the number of plaque-forming units (PFU) produced by FUdR-treated cells (10
3
to 10
4
PFU/ml) was at least 3 logs less than that produced by untreated cells, the yield of physical particles (as determined by electron microscopy) was approximately the same at 30 hr after infection. Besides being relatively non-infective, the particles produced in FUdR-treated cells appeared morphologically incomplete as they contained little or no nucleoid material.
Reovirus-induced Ribonucleic Acid Polymerase
Authors: Watanabe Y. , Gauntt C. J. , Graham A. F.
Published: January 2020

A virus-induced ribonucleic acid (RNA) polymerase activity was found in L cells infected with type 3 reovirus. Most of the enzyme is associated with the “large particle” fraction of the infected cells. The enzyme first appeared at 3 to 5 hr after infection and increased in amount until 7 to 9 hr. All four ribonucleoside triphosphates are incorporated in vitro into an acid-insoluble form by the enzyme. The major part of the product formed in vitro is a double-stranded RNA indistinguishable from viral RNA by electrophoresis on polyacrylamide gel. Approximately 40% of the product is a single-stranded RNA of relatively small molecular weight. More than 95% of the nucleotides incorporated into double-stranded RNA by the enzyme are bound in internal 3′-5′-phosphodiester linkages extending back from both 3′- and 5′-termini of the RNA strands.
Synthesis and Intracellular Localization of Vaccinia Virus Deoxyribonucleic Acid-dependent Ribonucleic Acid Polymerase
Authors: Kates J. , Dahl R. , Mielke M.
Published: January 2020

The time course of vaccinia deoxyribonucleic acid (DNA)-dependent ribonucleic acid (RNA) polymerase synthesis and its intracellular localization were studied with virus-infected HeLa cells. Viral RNA polymerase activity could be meassured shortly after viral infection in the cytoplasmic fraction of infected cells in vitro. However, unless the cells were broken in the presence of the nonionic detergent Triton-X-100, no significant synthesis of new RNA polymerase was detected during the viral growth cycle. When cells were broken in the presence of this detergent, extensive increases in viral RNA polymerase activity were observed late in the infection cycle. The onset of new RNA polymerase synthesis was dependent on prior viral DNA replication. Fluorodeoxyuridine (5 × 10
−5
m
) prevented the onset of viral polymerase synthesis. Streptovitacin A, a specific and complete inhibitor of protein synthesis in HeLa cells, prevented the synthesis of RNA polymerase. Thus, the synthesis of RNA polymerase is a “late” function of the virus. The newly synthesized RNA polymerase activity was primarily bound to particles which sedimented during high-speed centrifugation. These particles have been characterized by sucrose gradient centrifugation. A major class of active RNA polymerase particles were considerably “lighter” than whole virus in sucrose gradients. These particles were entirely resistant to the action of added pancreatic deoxyribonuclease, and they were not stimulated by added calf thymus primer DNA. It is concluded that these particles are not active in RNA synthesis in vivo, and that activation occurs as a result of detergent treatment in vitro.
Multiplication of parvovirus LuIII in a synchronized culture system. IV. Association of viral structural polypeptides with the host cell chromatin
Authors: Gautschi M , Siegl G , Kronauer G
Published: January 2020

Newly synthesized structural polypeptides of parvovirus LuIII, VP1 (62,000 daltons) and VP2 (74,000 daltons), were detected in nuclei of synchronized, infected HeLa cells at 11 to 12 h postinfection, i.e., after cells had passed through the S phase of the cell cycle. At this time, most of intranuclear viral polypeptides were associated with the chromatin acidic proteins. However, 13 to 14 h postinfection, about one-third of intranuclear VP1 and VP2 also could be extracted in the fraction containing nuclear sap proteins. According to pulse-chase experiments, VP1 and VP2 accumulated in the chromatin with a time lag of 20 to 30 min. About 90% of these chromatin-associated viral polypeptides represented empty viral capsids. In addition, chromatin prepared at 14 h postinfection contained 90 to 95% of the total intranuclear viral 16S replicative-form DNA. Since viral replicative-form DNA and empty viral capsids seem to be associated specifically with cellular chromatin, we assume that this subnuclear structure is the site of the synthesis of progeny viral DNA and the formation of complete virions.
Purification of the α Subunit of Avian Myeloblastosis Virus DNA Polymerase by Polyuridylic Acid-Sepharose
Authors: Grandgenett Duane P.
Published: January 2020

The α subunit of the avian myeloblastosis virus DNA polymerase could be readily purified to near homogeneity using a polyuridylic acid-Sepharose column chromatography step.
Transcription of the genome of adenovirus type 12. III. Maps of stable RNA from productively infected human cells and abortively infected and transformed hamster cells
Authors: Ortin J , Scheidtmann K H , Greenberg R , Westphal M , Doerfler W
Published: January 2020

The adenovirus type 12-specific mRNA and the stable nuclear RNA from productively infected KB cells, early postinfection, from abortively infected BHK-21 cells, and from the adenovirus type 12-transformed hamster lines T637 and HA12/7 have been mapped on the genome of adenovirus type 12. The intact separated heavy (H) and light (L) strands of adenovirus type 12 DNA have been used to determine the extent of complementarity of the mRNA or nuclear RNA from different cell lines to each of the strands. More precise map positions have been obtained by the use of the H and L complements of the fragments of adenovirus type 12 DNA which were produced with the EcoRI and BamHI restriction endonucleases. The results of the mapping experiments demonstrate that the mRNA's isolated early from productively and abortively infected and from two lines of transformed cells are derived from the same or similar regions of the adenovirus type 12 genome. The map positions on the adenovirus type 12 genome for the mRNA from the cell lines as indicated correspond to regions located approximately between 0 and 0.1 and 0.74 and 0.88 fractional length units on the L strand and to regions between 0.63 and 0.74 and 0.89 and 1.0 fractional length units on the H strand. The HA12/7 line lacks mRNA complementary to the region between 0.74 and 0.88 fractional length units on the L strand. Similar data are found for the nuclear RNA, except that the regions transcribed are more extensive than those observed in mRNA. The polarity of the H strand has its 3'-end on the right terminus in the EcoRI A fragment, and the L strand has its 3'-end on the left terminus in the EcoRI C fragment. Thus, the H strand is transcribed from right to left (1 = leftward strand); and the L strand is transcribed from left to right (r = rightward strand). The designations H and L refer to the relative heavy and light densities of the two strands in polyuridylic-polyguanylic acid-CsCl density gradients. The EcoRI C-H and D-H complements have been shown to be part of the intact L strand; thus, there is a "reversal in heaviness" on the left terminus of the viral DNA.
Mouse strain resistant to N-, B-, and NB-tropic murine leukemia viruses
Authors: Kai K , Ikeda H , Yuasa Y , Suzuki S , Odaka T
Published: January 2020

Mouse strain G was studied for its susceptibility to various strains of murine leukemia and sarcoma viruses. Both N- and NB-tropic Friend leukemia viruses neither induced splenomegaly nor grew efficiently in strain G mice. Using the XC test, cultured embryo cells were found to be resistant, but not absolutely, to all the tested viruses, N-tropic AKR virus, N- and NB-tropic Friend leukemia viruses, NB-tropic Rauscher leukemia virus, B-tropic WN1802B virus, NB-tropic Moloney leukemia and sarcoma viruses, and N-tropic Kirsten sarcoma virus, although the resistance to Moloney leukemia and sarcoma viruses is sometimes not as strong as that for other viruses. Thus, the strain G mice are unique among mouse strains because they show resistance that is not related to the N-B tropism of murine leukemia viruses.
Enzymatic basis for the selective inhibition of varicella-zoster virus by 5-halogenated analogues of deoxycytidine
Authors: Dobersen M J , Jerkofsky M , Greer S
Published: January 2020

5-Bromodeoxycytidine (BrdC) and 5-iododeoxycytidine, at a concentration of 100 mug/ml, effectively inhibit the replication of varicella-zoster (VZ) virus in tissue culture. No toxicity could be demonstrated in uninfected cells under the same conditions. Studies on the enzymatic basis for this selective inhibition were undertaken. Infection of human embryonic lung cell monolayers with VZ virus-infected cells results in the induction of thymidine (dT), deoxycytidine (dC), and BrdC kinase activities (which are increased 10-, 40-, and 60-fold, respectively) and in a 70-fold stimulation in the incorporation of 3H nucleotide (5-bromodeoxyuridylate) derived from BrdC into DNA. The thermal stability of the VZ virus-induced activities differs significantly from the activities induced by herpes simplex virus type 1 and herpes simplex virus type 2 and those present in uninfected human embryonic lung cells. The VZ virus-induced dT, dC, and BrdC kinase are similarly affected by temperature and cofractionate upon Sephadex gel filtration, findings consistent with the hypothesis that these activities are the function of a single enzyme: a pyrimidine deoxyribonucleoside kinase. The molecular weight, calculated on the basis of the elution pattern on Sephadex G-150, is 70,000. Kinetic studies, demonstrating that dT and dC competively inhibit the phosphorylation of BrdC, are consistent with the phosphorylation of these substrates at a common active site. Kinetic parameters include: KidT = 0.6 MUM; KidC = 60 muM; KmBrdC = 8.5 muM. In contrast to its relatively high affinity for the VZ virus-induced kinase, BrdC is a relatively poor substrate for the host kinases. Therefore, the basis for the selective inhibition of VZ virus by 5-halogenated analogues of dC is reflected in the induction of a pyrimidine deoxyribonucleoside kinase with a high affinity for BrdC.
Ultrastructural Studies of Hepatitis A Virus by Electron Microscopy
Authors: Cook E. H. , Bradley D. W. , Gravelle C. R. , Maynard J. E.
Published: January 2020

Well-defined, core-like structures were visualized in hepatitis A virus particles by a modified microelectron microscopy technique.
Initiation and Termination Mutants of
Bacillus subtilis
Bacteriophage SPO1
Authors: Glassberg Jeffrey , Franck Michelle , Stewart Charles R.
Published: January 2020

Mutants affected in cistrons 21 and 32 of bacteriophage SPO1 are defective specifically in the initiation of DNA replication. Mutations in cistron 32 also specifically affect the termination of replication.
Synthesis and processing of the precursor to the major core protein of adenovirus type 2
Authors: Everitt E , Meador S A , Levine A S
Published: January 2020

An isopycnic Metrizamide-detergent gradient system was developed in which the newly synthesized precursor (polypeptide P-VII) to the major core protein of adenovirus type 2 (polypeptide VII) was confined to a spectrum of complexes with densities equal to or higher than that of adenovirions. The majority of the newly synthesized P-VII was, at the beginning of the logarithmic period of virus production, present as an entity of protein density. This pool of P-VII was efficiently depleted. P-VII was also associated with high-molecular-weight structures of intermediate density, sharing some properties with empty capsids or incomplete particles. The transfer of P-VII from the intermediate-density region was not quantitative, and only particles of true virion density subsequently contained polypeptide VII. No structures equivalent to the core structure of disrupted virions or identical to incomplete particles were detected in this system. A temperature-dependent transition of radioactivity from polypeptide P-VII into polypeptide VII was also detectable after in vitro incubation of P-VII-containing complexes. Addition of Ad2-infected cell extracts was required for processing of complexes derived from regions of protein density, whereas P-VII was processed spontaneously upon incubation in complexes of virion density.
Role of the pneumococcal autolysin (murein hydrolase) in the release of progeny bacteriophage and in the bacteriophage-induced lysis of the host cells
Authors: Ronda-Lain C , Lopez R , Tapia A , Tomasz A
Published: January 2020

The pneumococcal bacteriophage Dp-1 seems to require the activity of the N-acetylmuramic acid-L-alanine amidase of the host bacterium for the liberation of phage progeny into the medium. This conclusion is based on a series of observations indicating that the exit of progeny phage particles is prevented by conditions that specifically inhibit the activity of the pneumococcal autolysin. These inhibitory conditions are as follows: (i) growth of the bacteria on ethanolamine-containing medium; (ii) growth of the cells at pH values that inhibit penicillin-induced lysis of pneumococcal cultures and lysis in the stationary phase of growth; (iii) addition of trypsin or the autolysin-inhibitory pneumococcal Forssman antigen (lipoteichoric acid) to the growth medium before lysis; (iv) infection of an autolysin-defective pneumococcal mutant at a multiplicity of infection less than 10 (treatment of such infected mutant bacteria with wild-type autolysin from without can liberate the entrapped progeny phage particles); (v) release of phage particles and culture lysis can also be inhibited by the addition of chloramphenicol to infected cultures just before the time at which lysis would normally occur. Bacteria infected with Dp-1 under conditions nonpermissive for culture lysis and phage release secrete into the growth medium a substantial portion of their cellular Forssman antigen in the form of a macromolecular complex that has autolysin-inhibitory activity. We suggest that a phage product may trigger the bacterial autolysin by a mechanism similar to that operating during treatment of pneumococci with penicillin (Tomasz and Waks, 1975).
Polypeptides of respiratory syncytial virus
Authors: Levine S
Published: January 2020

Radiolabeled respiratory syncytial virus was purified from medium that had been harvested from infected HeLa cell monolayers before it contained much cellular debris. After isopycnic centrifugation in linear gradients prepared with sucrose dissolved in Hanks balanced salt solution, almost all the infectivity and most of the radioactivity were recovered in a single band with density from 1.16 to 1.23 g/cm3 and a peak at 1.2 g/cm3. Analysis by polyacrylamide gel electrophoresis resolved the purified virus into seven polypeptides of approximate molecular weights 20,000 to 80,000, of which the two largest and the smallest proved to by glycoproteins.