bluuebunny/crossref_metadata_2025 huggingface dataset:
Thymidine kinase not required for antiviral activity of acyclovir against mouse cytomegalovirus
Authors: Burns W H , Wingard J R , Bender W J , Saral R
Published: January 2020

Previous studies of herpesvirus infections have indicated that a virus-specified thymidine kinase is required for the initial phosphorylation of acyclovir [acycloguanosine or 9-(2-hydroxyethoxymethyl)guanine] in the formation of acycloguanosine triphosphate. The latter compound accumulates in infected cells and competitively inhibits the viral DNA polymerase. We found that mouse cytomegalovirus, which does not express a thymidine kinase, was sensitive to the antiviral effects of acyclovir at a 50% inhibitory dose of approximately 0.23 microM. Acyclovir was equally effective against mouse cytomegalovirus in normal 3T3 cells and in 3T3 cells deficient in cellular thymidine kinase. Furthermore, the activity of acyclovir could not be reversed by excess thymidine, which easily reversed the antiviral activity of acyclovir against herpes simplex virus. Using a high-pressure liquid chromatography technique that easily detected acycloguanosine triphosphate in cells infected with herpes simplex virus, we could not detect acycloguanosine triphosphate in mouse cytomegalovirus-infected cells. These experiments demonstrated that the activity of acyclovir against mouse cytomegalovirus is not dependent on a thymidine phosphorylation pathway. Additional experiments are underway to determine whether acycloguanosine triphosphate is produced by another pathway in concentrations sufficient to inhibit mouse cytomegalovirus DNA polymerase.
Early Events in Herpes Simplex Virus Infection: a Radioautographic Study
Authors: Hummeler Klaus , Tomassini Natale , Zajac Barbara
Published: January 2020

The early events in herpes simplex virus infection were studied by means of radio-autography. The virus was rapidly taken up by the host cells and uncoated. Viral deoxyribonucleic acid (DNA) reached the nuclear sites of replication in 15 to 30 min after infection. The viral DNA occasionally associated with chromosomes or condensed chromatin but was more frequently found to be randomly distributed. Viral progeny appeared 3 hr after infection. These particles did not show any particular spatial relationship to the parental DNA. The morphological latent period lasted 2.5 hr.
Replication of Canine Herpesvirus
Authors: Aurelian Laure
Published: January 2020

This paper reports the results of two series of experiments. The first series indicated that deoxyribonucleic acid (DNA) extracted from partially purified canine herpesvirus virions is characterized by a high guanine plus cytosine molar base ratio (65 to 67 mole%), similar to the DNA of herpes simplex virus. In the second series of experiments it was estimated, on the basis of uptake of tritiated thymidine, that in dog kidney cells canine herpesvirus-DNA synthesis starts at 4 hr and continues until 16 hr after infection. Treatment of infected cells with puromycin during the first 4 hr of infection blocks the onset of viral DNA synthesis, whereas, after this time the uptake of thymidine is unaffected.
Delayed Lysis with
Salmonella
Bacteriophage P22: Induction of Lysis by Addition of Cysteine or Histidine to the Growth Medium
Authors: Cohen Larry W.
Published: January 2020

A mutant (
Lys
−
) of
Salmonella
bacteriophage P22 showed a delay in lysis of more than 3 hr in infections in unsupplemented M9 medium. The infected cells were induced to lyse during that interval by addition of histidine or sulfhydryl compounds cysteine, mercaptoethanol, glutathione, or ergothioneine. Urocanic acid, the first intermediate in the catabolic histidine pathway, did not induce lysis, nor did histamine, imidazolelactate, or carnosine. None of the other amino acids common to protein had any inductive effect. Both the
d
and
l
forms of histidine were effective in inducing lysis, suggesting that the incorporation of the histidine into protein is not involved. Chloramphenicol inhibited lysis when added at 60 min with or without histidine, but did not inhibit the induction of lysis when added with cysteine. Bacterial cells infected with
Lys
+
phage were induced to lyse prematurely when cysteine was added at 30 min but not at 20 min of infection. Iodoacetate inhibited lysis of
Lys
+
-infected cells when added at 20 min but not at 30 min.
Cells Persistently Infected with Newcastle Disease Virus
Authors: Thacore Harshad , Youngner Julius S.
Published: January 2020

The strain of Newcastle disease virus (NDV
pi
) present in persistently infected L cells differed markedly from the Herts strain (NDV
0
) used to initiate the infection. NDV
pi
produced small plaques (less than 1 mm) in chick embryo cell cultures, whereas the wild type (NDV
0
) produced large plaques (2 to 3 mm). The two viruses differed in a number of additional properties. Whereas 80% of adsorbed NDV
0
eluted from chicken red blood cells at 37 C, only about 20% of NDV
pi
was recovered under similar conditions. There was no significant difference in the neuraminidase content of the two viruses. The infectivity of NDV
0
was stable for 1 hr at 48 C, whereas 99.9% of the infectivity of NDV
pi
was destroyed. The two viruses also differed in lethality for chick embryos; NDV
pi
had significantly reduced lethality for 9-day-old chick embryos when compared to NDV
0
. In contrast to NDV
0
, which produced an abortive infection in L cells, NDV
pi
not only replicated effectively and destroyed these cells, but also induced significantly higher quantities of interferon than did NDV
0
. These data furnished additional evidence for the lack of relationship of interferon production to abortive infection of L cells with NDV
0
. In contrast, interferon was found to play a significant role in the maintenance of persistent infection.
Phenotypic Suppression by Streptomycin of Amber Mutants in the Ribonucleic Acid Bacteriophage Coat Protein Cistron
Authors: Kuwano Michihiko , Endo Hideya
Published: January 2020

After mutagenesis with nitrosoguanidine or ultraviolet light, 298 streptomycin high-resistant and 98 streptomycin high-dependent mutants were isolated from HfrC Su
−
. They were tested for their ability to phenotypically suppress five different amber ribonucleic acid (RNA) bacteriophage mutants in the presence of streptomycin. The phage mutants are all in the coat protein, which is 129 amino acids long; the uracil-adenine-guanine codons were at the following positions:
sus
3 and
am
B2, 6;
am
B11, 50;
am
B21, 54;
sus
11, 70. Only
sus
3 and
am
B2 could be phenotypically suppressed by streptomycin; this was clearly demonstrated in nine mutant strains, seven
str
-HR and two
str
-HD. The suppression was always dependent upon added streptomycin and was dose-dependent in all cases. None of the mutants showed measurable suppression in absence of the drug. Among revertants to streptomycin independence from streptomycin-dependent strains that could show phenotypic suppression, most of those that were still resistant to streptomycin (10 μg or more) retained the capacity to show phenotypic suppression; whereas among those revertants sensitive to 10 μg of streptomycin or less, none retained the capacity. Eight different amber polar mutants (strong and weak) in gene 34 of phage T4 were also tested for pleiotypic suppression by streptomycin in all the streptomycin-resistant and -dependent strains isolated. No suppression was found in any of the 396 strains tested.
Fate of Adenovirus Type 12 Genomes in Nonpermissive Cells
Authors: zur Hausen Harald , Sokol Frantisek
Published: January 2020

The fate of
3
H-thymidine-labeled adenovirus type 12 deoxyribonucleic acid (DNA) was studied in Nil-2 cells of Syrian hamster origin. It was found that a substantial fraction of
3
H-adenovirus type 12 DNA became degraded within 24 hr after infection and was released into the culture fluid. After infection of 5-bromodeoxyuridine (BUdR)-prelabeled cells with
3
H-adenovirus type 12, viral DNA became readily separable from cellular DNA by equilibrium centrifugation in CsCl. Part of the viral radioactivity was found to shift gradually to the position of cellular DNA as time progressed after infection. When exponentially growing cells were exposed simultaneously to BUdR, 5-fluorodeoxyuridine, and
3
H-adenovirus type 12, up to 50% of the viral radioactivity shifted within 24 hr from the density of viral DNA to that of cellular DNA after equilibrium centrifugation in CsCl. Upon denaturation of the cellular DNA, the isotope was preferentially found to be associated with the “heavy” strand which was synthesized after infection. Upon hybridization of the “heavy” and the “light” strands with sonically treated, denatured
3
H-adenovirus type 12 DNA, small and nearly equal amounts of counts hybridized with both strands. The number of counts annealed was in a range similar to that of those annealed with the same amount of DNA derived from adenovirus type 12-transformed hamster cells. These results demonstrate that (i) a substantial proportion of the adsorbed virus becomes degraded within 24 hr; (ii) part of the degradation products is reutilized for cellular DNA synthesis; (iii) only a small fraction, mainly fragments, of viral DNA becomes integrated into both the newly synthesized and the parental strands of cellular DNA.
Precursor Products Found in Formaldehyde-fixed Lysates of BHK-21 Cells Infected with Pseudorabies Virus
Authors: Sydiskis Robert J.
Published: January 2020

Lysates of BHK-21 cells, infected with pseudorabies and labeled with
3
H-thymidine, were treated with formaldehyde and centrifuged in preformed CsCl gradients. Five peaks, designated bands A to E, were detected and shown to contain the following virus-specific products: band A, aggregates of virions; band B, virions; band C, empty capsids and nucleocapsids; bands D and E, small nucleoid-like particles. All five bands contained viral deoxyribonucleic acid and virous-specific antigens. Deoxyribonuclease treatment of the lysates before centrifugation resulted in the complete loss of
3
H-thymidine-labeled material from bands D and E; bands B and C were unaffected. Pulse-chase studies showed that
3
H-thymidine can be rapidly chased into the virus-specific products sedimenting in bands D and E. Further chase resulted in the gradual loss of label from bands D and E with a concomitant increase in the amount of label in bands A, B, and C. These results indicate that the virus-specific products in bands D and E are precursors of pseudorabies virions and may represent the nucleoid of the virion.
Biophysical Properties of Frog Virus and Its Deoxyribonucleic Acid: Fate of Radioactive Virus in the Early Stage of Infection
Authors: Smith William R. , McAuslan Brian R.
Published: January 2020

Frog virus (FV-3) was banded by isopycnic centrifugation in cesium chloride, sucrose, or potassium tartrate. Two bands of infectivity were regularly found at positions in cesium chloride corresponding to densities of 1.26 and 1.30 g/cm
3
, respectively. Deoxyribonucleic acid from either band had the following characteristics: double-stranded; a T
m
of 76.3 C in 0.1 SSC (0.015
m
NaCl plus 0.015
m
sodium citrate) and a buoyant density of 1.720 g/cm
3
in cesium chloride, corresponding to a guanine plus cytosine content of 56 to 58% and a molecular weight of 130 × 10
6
daltons, determined by velocity sedimentation. These data, together with electron micrographs of sections of cells infected with material from either band suggest that two types of infectious frog virus particles exists, rather than a second virus in the frog virus stocks. The composition of frog virus was determined. It was found that highly purified preparations of frog virus were composed of 55.8% protein, 30.1% deoxyribonucleic acid, and 14.2% lipid. The kinetics of adsorption and uncoating of FV-3 was studied with radioactive virus. Uncoating is comparatively rapid and in contrast to poxvirus is unaffected by inhibitors of protein synthesis.
Proteins of Newcastle Disease Virus and of the Viral Nucleocapsid
Authors: Bikel Ilan , Duesberg Peter H.
Published: January 2020

Newcastle disease virus was found to contain three major proteins. The structure unit of the viral nucleocapsid appears to be monomeric and to consist of a single large protein of an approximate molecular weight of 62,000.
Biological Effects of Substituting Cytosine for 5-Hydroxymethylcytosine in the Deoxyribonucleic Acid of Bacteriophage T4
Authors: Kutter Elizabeth M. , Wiberg John S.
Published: January 2020

Previous work from this laboratory has shown that the cytosine-containing T4 deoxyribonucleic acid (DNA) made by deoxycytidine triphosphatase (dCTPase)
amber
mutants is extensively degraded, and that nucleases controlled by genes 46 and 47 participate in this process. In this paper, we examine other consequences of a defective dCTPase. Included are studies of DNA synthesis and phage production, and of the control of both early and late protein synthesis after infection of
Escherichia coli
B with various T4 mutants defective in genes 56 (dCTPase), 42 (dCMP hydroxymethylase), 1 (deoxynucleotide kinase), 43 (DNA polymerase), 30 (polynucleotide ligase), 46 and 47 (DNA breakdown) or
e
(lysozyme). By varying the temperature of infection with a temperature-sensitive dCTPase mutant, we have been able to control intracellular dCTPase activity, and thus vary the cytosine content of the phage DNA. We have produced and characterized viable T4 phage in which cytosine replaces 20% of the 5-hydroxymethylcytosine (HMC) in the DNA. We present evidence which suggests that intact, cytosine-containing T4 DNA is much less efficient than is normal T4 DNA in directing the synthesis of tail-fiber antigen. Lysozyme production is much less affected by progressively decreasing dCTPase activity; however, complete substitution of cytosine is correlated with a depression of lysozyme synthesis greater than expected from the defective synthesis of DNA. Low but significant lysozyme synthesis is observed late after infection of
E. coli
B with T4
amber
mutants defective in a number of genes controlling DNA synthesis. The “20% cytosine” T4 phage, once produced, can initiate an apparently normal infection at permissive temperatures; the synthesis of early enzymes, DNA, and phage does not appear to be impaired. Two roles for HMC in T4 DNA have been indicated previously: (i) involvement in host-controlled restriction of the phage, in which glucosylation of the hydroxymethyl group plays a crucial role (16, 29, 53, 58), and (ii) protection of vegetative DNA against phage-controlled nucleases, a protection not dependent on glucosylation (41, 66, 67). A third role is suggested by our present results: transcription of at least some late genes can occur only from HMC-containing DNA and not from cytosine-containing DNA.
Serological Relatedness of the Ribonucleic Acid-Containing Coliphages
Authors: Krueger Robert G.
Published: January 2020

The serological relationship of the ribonucleic acid (RNA)-containing coliphages MS-2, M-12, R-17, f
2
, β, fr, f
4
, and Qβ was determined. Antisera against MS-2, R-17, f
2
, fr, and Qβ neutralized the infectivity of all of these RNA phages to varying degrees. Although each phage was serologically distinct, the antisera cross-reacted with certain phages to approximately the same degree, indicating the antigenic relationship of the coat proteins of these phages. Adsorption of anti-MS-2 sera with varying concentrations of all of the phages demonstrated that these viruses contain similar yet unique antigenic determinants. It is suggested that these RNA phages are mutants of two related phages rather than of the same phage.
In Vitro Transformation by Adenovirus-Simian Virus 40 Hybrid Viruses
Authors: Black Paul H. , Berman Leonard D. , Dixon Cynthia B.
Published: January 2020

Clones were isolated from hamster cells transformed by the adenovirus 2-SV40 and adenovirus 12-SV40 transcapsidant hybrid viruses. The clones were characterized with respect to their cytomorphology, virus and antigen content, and the histomorphology of tumors induced by transplantation of the clonal sublines to hamsters. Three different cellular and colonial morphologies were observed. Clones with an SV40 morphology gave rise to tumors predominantly with an SV40 histology, whereas clones with an adenovirus morphology produced typical adenovirus tumors upon transplantation of the transformed cells. Clones which had features of both SV40 and adenovirus transformed cells gave rise to “intermediate” and adenovirus tumors. The results indicate that multiple events occur during transformation and tumorigenesis by the transcapsidant virus populations and provide an explanation for the multiplicity of findings which have been reported with these virus populations.
Buoyant Density Analysis of Staphylococcal Bacteriophage 80 Transducing Particles
Authors: Berryhill David L. , Pattee P. A.
Published: January 2020

Densities of
pase, tet
, and
ol
transducing particles were indistinguishable, and about 0.002 g × cm
−3
less dense than the plaque-forming units (PFU). The densities of the PFU and
nov
transducing particles were indistinguishable, the PFU having a density of 1.507 g × cm
−3
.
Endogenous type C retroviral sequences of mice are organized in a small number of virus-like classes and have been acquired recently
Authors: Dolberg D S , Bacheler L T , Fan H
Published: January 2020

We studied endogenous type C virus-related sequences of mice by annealing Moloney murine leukemia virus DNA to agarose gel blot transfers of uninfected mouse cell DNA which had been cleaved with restriction enzymes. We found that many of the endogenous murine leukemia virus-related sequences in mice consist of two organizational classes that are integrated into many different loci. Both of these classes resemble standard murine leukemia virus proviral DNA in both size and sequence organization. All lines of inbred mice examined contained both organizational classes, as did feral isolates of Mus musculus domesticus. However, a related Asian subspecies, Mus musculus molossinus, contained different organizational classes of endogenous murine leukemia virus-related sequences. Among inbred strains and feral isolates of M. musculus domesticus, the murine leukemia virus-related sequences were present at different loci. This suggested that most of these sequences were acquired relatively recently during subspeciation and inbreeding.
The transient inability of the conjugating female cell to host 186 infection explains the absence of zygotic induction for 186
Authors: Woods W H , Egan B
Published: January 2020

In an Hfr(186) X F- cross, the 186 prophage on the incoming male chromosome is not induced, despite the fact that prophage 186 can be induced by other means (W. H. Woods and J.B. Egan, J Virol. 14:1349-1356, 1974). We show here that the conjugating female is temporarily inhibitory to infection by 186, and this delay, we postulate, enables cI repression to be reestablished before the female cell recovers its 186 sensitivity.
Control of adenovirus early gene expression: accumulation of viral mRNA after infection of transformed cells
Authors: Persson H , Katze M G , Philipson L
Published: January 2020

We studied the accumulation of viral mRNA in the presence of inhibitors of protein synthesis in an adenovirus type 5-transformed cell line (line 293 cells). An analysis of the endogenous viral mRNA's and proteins revealed that only early regions 1A and 1B were expressed in uninfected 293 cells. However, viral mRNA's from early regions 2, 3, and 4, as well as mRNA's from early regions 1A and 1B, accumulated in 293 cells after infection with adenovirus type 2. Cells treated with anisomycin before infection showed a drastic enhancement of mRNA from early region 4 compared with drug-free controls, This increase in viral mRNA was detected by using filter hybridization, S1 endonuclease mapping, and in vitro translation. The rate of transcription of early region 4 nuclear RNA also increased significantly in the presence of anisomycin. In contrast to the results with early region 4, the levels of cytoplasmic mRNA's from early regions 2 and 3 did not increase in cells treated with inhibitors. These results suggested that multiple virus encoded controls operate on the early regions of the adenovirus genome.
Excision repair and patch size in UV-irradiated bacteriophage T4
Authors: Yarosh D B , Rosenstein B S , Setlow R B
Published: January 2020

We determined the average size of excision repair patches in repair of UV lesions in bacteriophage T4 by measuring the photolysis of bromodeoxyuridine incorporated during repair. The average patch was small, approximately four nucleotides long. In control experiments with the denV1 excision-deficient mutant, we encountered an artifact, a protein(s) which remained bound to phenol-extracted DNA and prevented nicking by the UV-specific endonucleases of Micrococcus luteus and bacteriophage T4.
Herpes simplex virus type 1 and 2 intracellular p40: type-specific and cross-reactive antigenic determinants on peptides generated by partial proteolysis
Authors: Heilman C J , Zweig M , Hampar B
Published: January 2020

Intracellular p40 is a class of protein ranging in molecular weight from 39,000 to 45,000 that is immunoprecipitated from herpes simplex virus type 1 (HSV-1)- and HSV-2-infected cell extracts by mouse monoclonal antibodies or guinea pig antisera against HSV-1 and HSV-2 nucleocapsid p40. Analysis by a two-dimensional gel system showed that HSV-1 and HSV-2 intracellular p40 each consisted of three major components. However, these HSV-1 and HSV-2 proteins differed in charge and size. Analysis of Staphylococcus aureus V8 protease partial digests by two-dimensional gel electrophoresis indicated that none of the peptides of HSV-1 and HSV-2 intracellular p40 were identical. Immunoprecipitation of the partial digest products of intracellular p40-1 and p40-2 with homologous and heterologous guinea pig antisera resulted in the precipitation of various combinations of peptides indicating the presence of either type-specific or cross-reactive antigenic determinants.
Immunological and Chemical Identification of Intracellular Forms of Adenovirus Type 2 Terminal Protein
Authors: Green Maurice , Symington Janey , Brackmann Karl H. , Cartas Maria A. , Thornton Helen , Young Leann
Published: January 2020

Highly purified adenovirus type 2 terminal protein (TP) with an apparent
M
r
of 55,000 (55K) was prepared in quantities of 10 to 30 μg from guanidine hydrochloride- or sodium dodecyl sulfate-disrupted virions (60 to 120 mg). Guinea pigs were immunized with 14 to 20 injections of TP in amounts of 1 to 2 μg. Antiserum to TP was used to study the intracellular polypeptides related to adenovirus type 2 TP. By immunoprecipitation with anti-TP serum, we identified 80K and 76K polypeptides in the nucleoplasmic and cytoplasmic S100 fractions of [
35
S]methionine-labeled cells early and late after infection with Ad2. By immunoautoradiographic analysis which eliminates coprecipitation of unrelated proteins, we identified an 80K polypeptide (probably an 80K-76K doublet) in unlabeled, late infected cells, using anti-TP serum and
125
I-labeled staphylococcal protein A. About two- to threefold-higher levels of the 80K and 76K polypeptides were present in the nucleoplasm than in the S100 fraction, and two- to threefold-higher levels were found in late infected cells than in early infected cells (cycloheximide enhanced, arabinofuranosylcytosine treated). We did not detect the 80K or 76K polypeptide in uninfected cells, indicating that these polypeptides are virus coded. Tryptic peptide map analysis showed that the 80K and 76K polypeptides are very closely related and that they share peptides with the DNA-bound 55K TP. Our data provide the first direct demonstration of intracellular 80K and 76K forms of TP. The intracellular 80K and 76K polypeptides are closely related or identical to the 80K polypeptide that Challberg and co-workers (Proc. Natl. Acad. Sci. U.S.A. 77:5105-5109, 1980) detected at the termini of adenovirus DNA synthesized in vitro and to the 87K polypeptide that Stillman and co-workers (Cell 23:497-508, 1981) translated in vitro. We did not detect the 55K TP in early or late infected cells, consistent with the proposal by Challberg and co-workers that the 80K polypeptide is a precursor to the virion-bound TP and that the conversion of the 80K polypeptide to the 55K TP occurs during virus maturation. The 80K and 76K polypeptides have many more methionine-containing tryptic peptides than does the 55K TP, and most of the tryptic peptides unique to the 80K and 76K polypeptides are very hydrophobic. Thus, the conversion of the 80K and 76K polypeptides to the 55K TP may involve the removal of a specific hydrophobic protein region.
Expression of adenovirus type 2 late genes in HeLa cells infected with adenovirus type 2 or adenovirus type 2-simian virus 40 hybrid viruses
Authors: Lawrence C B
Published: January 2020

The relative levels of expression of the five 3'-coterminal mRNA families encode in the 16.4 to 99 map unit transcription unit of adenovirus type 2 (Ad2) are determined in part by the frequency of utilization of five possible polyadenylation sites. The possibility that polyadenylation frequencies at these sites may be regulated was tested by examining the expression of the fiber (polypeptide IV) gene in cells infected with nondefective (ND) Ad2-simian virus 40 hybrid viruses which express a sixth late mRNA family. Both the relative rate of fiber protein synthesis and the relative abundance of fiber mRNA were normal in cells infected with the hybrid viruses Ad2+ND1, Ad2+ND2, or Ad2+ND4.
Relationships Among Virus Spread, Cytopathogenicity, and Virulence as Revealed by the Noncytopathic Mutants of Newcastle Disease Virus
Authors: Madansky Charles H. , Bratt Michael A.
Published: January 2020

We have studied protein synthesis in cultured cells infected with the six noncytopathic (
nc
) mutants of the Australia-Victoria strain (AV-WT) of Newcastle disease virus and their plaque-forming revertants. Virus-specific polypeptides accumulated at 30 to 63% of wild-type levels in
nc
mutant-infected cells and between 66 and 175% of wild-type levels in revertant-infected cells. An exception was the L polypeptide, which accumulated in
nc
mutant-infected cells at only 5 to 20% of the levels found in wild-type infection. The reduced accumulation of the L polypeptide did not appear to be due to increased degradation of that polypeptide. A new polypeptide (X) accumulated instead of polypeptide P in cells infected with mutants
nc
4 or
nc
16 and in virions released from them. Peptide mapping identified X as an altered form of P. A revertant of mutant
nc
4 (
nc
4S1), which forms larger hemadsorbing spots, but still does not form plaques, accumulated P instead of the X polypeptide. Thus, a lesion in P can affect virus spread without affecting cytopathogenicity. Virions of mutant
nc
7 and two naturally occurring avirulent strains of Newcastle disease virus (NJ LaSota and B1-Hitchner) contained polypeptides (F
7
and F
A
, respectively) related to, but migrating more rapidly than, F
0
in sodium dodecyl sulfate-polyacrylamide gels. As previously reported for avirulent strains, a brief treatment of
nc
7 virions with trypsin converted F
7
to F and increased infectivity. Similarly, culturing
nc
7-infected cells in the presence of trypsin facilitated fusion from within and viral spread from cell to cell. A plaque-forming revertant of
nc
7 still accumulated F
7
in virions, indicating that the lesions responsible for the F
7
and noncytopathic phenotypes are genetically separable. The virulent parental strain, AV-WT, exhibited a mean embryo death time of 42 h. Both the larger-spot-forming revertant of
nc
4 (
nc
4S1) and the small-plaque-forming revertant of
nc
7 exhibited a decrease in mean embryo death time (increase in virulence) from 74 to 63 h. A second-step, plaque-forming revertant derived from
nc
4S1 (
nc
4S1R1) exhibited a further decrease in mean embryo death time from 63 to 44 h. The results suggest that the F
A
-F
7
and X lesions affect the ability of virus to spread from cell to cell. In addition, these lesions appear to be genetically separable from those responsible for the noncytopathic phenotype. However, both types of lesions cause an extension of mean embryo death time and, thus, may be relevant to virulence in vivo.
Hybridization Selection and In Vitro Translation of
Autographa californica
Nuclear Polyhedrosis Virus mRNA
Authors: Vlak Just M. , Smith Gale E. , Summers Max D.
Published: January 2020

We isolated polyadenylated RNA from the cytoplasm of cells infected with
Autographa californica
nuclear polyhedrosis virus late after infection (21 h postinfection). At that time intracellular protein synthesis was directed almost exclusively toward infected cell-specific proteins. The polyadenylic acid-containing RNA sequences in the cytoplasm at 21 h postinfection were radiolabeled in vitro and hybridized to
A. californica
nuclear polyhedrosis virus DNA restriction fragments. The polyadenylic acid-containing RNA was derived from regions representing the entire viral genome. Translation in a reticulocyte cell-free protein-synthesizing system of cytoplasmic RNA selected by hybridization to viral DNA and polyadenylic acid-containing RNA produced almost identical polypeptide patterns, suggesting that late after infection almost all of the cytoplasmic polyadenylic acid-containing RNA present in infected cells was of viral origin. Polyhedrin protein (molecular weight, 33,000) and a number of virion structural proteins were among the translation products which were identified by immunoprecipitation and by comparing molecular weights. In addition, some tentative nonstructural infected cell-specific proteins were also detected. Using the hybridization selection technique, we determined that sequences complementary to the message coding for polyhedrin were located on
Eco
RI fragment I of
A. californica
nuclear polyhedrosis virus DNA, whereas sequences coding for a putative nonstructural protein (molecular weight, 39,000) were on
Eco
RI fragment J.
Properties of an intracisternal A-particle-associated endonuclease activity which is stimulated by ATP
Authors: Nissen-Meyer J , Eikhom T S
Published: January 2020

An endonuclease activity associated with purified proteinase K-treated intracisternal A-particles was identified and characterized. The activity required divalent cations, preferring Mn2+ to Mg2+. Salt concentrations above 50 mM inhibited the activity. The endonuclease was greatly stimulated by ATP, ADP, and dATP, whereas AMP appeared to produce a slight inhibition. GTP had no apparent effect on the activity. The enzyme introduced single-stranded nicks into DNA and nicked preferentially supercoiled DNA duplexes in the presence of ATP, although linear duplexes also functioned as substrates. Single-stranded DNA was not nicked to any great extent. The molecular weight of the enzyme was estimated to be about 40,000. The characteristics of this enzyme are very similar to those of the endonuclease found associated with Friend murine leukemia virus.
Characterization of N-type and dually permissive cells segregated from mouse fibroblasts whose Fv-1 phenotype could be modified by another independently segregating gene(s)
Authors: Yoshikura H , Tejima S , Kuchino T , Segawa K , Odaka T
Published: January 2020

Though the inbred DDD mouse strain is essentially of the N type, the primary culture of this strain was about 100-fold more sensitive to B-tropic WN1802B virus than were the typical N-type strains (C3H/He, C57L, etc.). After cloning, DDD mouse cells segregated two types of cells, typical N-type cells and cells lacking in Fv-1 restriction. As both types of cells so far tested retained glucose-6-phosphatase-1 coded by a locus closely linked to Fv-1 and genetic cross experiments indicated the presence of a gene(s) modifying the Fv-1 phenotype, variation in Fv-1 restriction could presumably be brought about by genetic changes in a gene(s) other than Fv-1 itself. N-type and dually permissive cell clones were similarly established from the inbred G mouse. Compositions of polypeptides labeled with [35S]methionine in the N-type and dually permissive cells of DDD and G mouse origins were compared by two-dimensional gel electrophoresis. The polypeptide maps of these cells were similar except for a few spots. Among these dissimilar spots, a spot of about 20,000 daltons with a pI of about 5.5 was always present in N-type cells, whereas it was absent in dually permissive cells. In DDD mouse-derived clones, a proportional relation was observed between the intensity of the spot and the restriction to the B-tropic virus.
Establishment of infection by spleen necrosis virus: inhibition in stationary cells and the role of secondary infection
Authors: Chen I S , Temin H M
Published: January 2020

The relationship of two early events in the establishment of infection by avian retroviruses, the inhibition of viral DNA synthesis in stationary avian cells and the secondary infection which occurs after infection of replicating cells, was investigated. When neutralizing antibody to spleen necrosis virus was used to prevent secondary infection, the amount of unintegrated linear spleen necrosis virus DNA detected was much lower in infected stationary cells than in infected replicating cells. The amount of unintegrated linear spleen necrosis virus DNA in stationary cells was less than one copy per cell even at high multiplicities of infection. Viral DNA synthesis resumed after stimulation of the cells to replicate. The time of this viral DNA synthesis was closely correlated with renewed cellular DNA synthesis. In addition, blocking secondary infection of replicating cells prevented the rate of virus production from reaching the high levels usually associated with a normal productive infection by SNV. Virus production increased if secondary infection was allowed. However, this rise in virus production was not proportional to the amounts of viral DNA integrated after secondary infection.
Integration site of polyoma virus DNA in the inducible LPT line of polyoma-transformed rat cells
Authors: Mendelsohn E , Baran N , Neer A , Manor H
Published: January 2020

The structure of the polyoma virus (Py) integration site in the inducible LPT line of Py-transformed rat cells was determined by biochemical methods of gene mapping. LPT cell DNA was digested with various restriction enzymes. The digestion products were electrophoresed in agarose gels and transferred onto nitrocellulose sheets by Southern blotting. Fragments containing viral or cell DNA sequences, or both, were identified by hybridization with Py DNA or with a cloned flanking cell DNA probe. Cleavage of LPT DNA with enzymes that restrict the Py genome once generated linear Py DNA molecules and two fragments containing both cell and viral DNA sequences. Cleavage of LPT DNA with enzymes which do not restrict Py DNA generated series of fragments whose lengths were found to differ by increments of a whole Py genome; the smallest fragment in each series was found to be longer than the viral genome. These data indicate that LPT cultures contain Py insertions of various lengths integrated into the same chromosomal site in all the cells. The length heterogeneity of the viral insertions is due to the presence of 0, 1, 2, 3. . . Py genomes arranged in a direct tandem repeat within invariable sequences of viral DNA. Double-digestion experiments were also carried out with the above enzymes and with enzymes that cleave the Py genome at multiple sites. The data obtained in these experiments were used to construct a physical map of the integration site. This map showed that the early region of the virus remained intact even in the smallest insertion (which contains no whole duplicated genomes), whereas the late region was partially duplicated and split during integration. The smallest insertion is colinear with the Py physical map over a region including the entire Py genome and at least a part of the duplicated segment. This structure could give rise to nondefective circular viral DNA molecules by single homologous recombination events. Similar recombination events may occur at a higher frequency in the longer insertions, which include longer regions of homology, and may yield many more free viral genomes. The presence of these insertions in LPT cells could thus be one of the factors which account for the high inducibility of the LPT line.
Evidence for intramolecular self-cleavage of picornaviral replicase precursors
Authors: Palmenberg A C , Rueckert R R
Published: January 2020

It has previously been shown that when encephalomyocarditis viral RNA is translated in cell-free extracts of rabbit reticulocytes, it synthesizes a virus-coded protease, p22, which is derived by cleavage of a precursor protein, C. Protein C is shown here to be cleaved by two different mechanisms, which were distinguished by their sensitivity to dilution. One mechanism was sensitive to dilution; the other was not. The biphasic cleavage behavior was unchanged by diluting incubation mixtures with untranslated reticulocyte extract instead of buffer, suggesting that both types of cleavage were mediated by virus translation products. It is proposed that the dilution-sensitive cleavage of protein C is due to a virus-coded protease, probably p22 itself, and that the dilution-independent cleavage is due to intramolecular self-cleavage of protein C.
Cleavage sites within the poliovirus capsid protein precursors
Authors: Larsen G R , Anderson C W , Dorner A J , Semler B L , Wimmer E
Published: January 2020

Partial amino-terminal sequence analysis was performed on radiolabeled polio-virus capsid proteins VP1, VP2, and VP3. A computer-assisted comparison of the amino acid sequences obtained with that predicted by the nucleotide sequence of the poliovirus genome allows assignment of the amino terminus of each capsid protein to a unique position within the virus polyprotein. Sequence analysis of trypsin-digested VP4, which has a blocked amino terminus, demonstrates that VP4 is encoded at or very near to the amino terminus of the polyprotein. The gene order of the capsid proteins is VP4-VP2-VP3-VP1. Cleavage of VP0 to VP4 and VP2 is shown to occur between asparagine and serine, whereas the cleavages that separate VP2/VP3 and VP3/VP1 occur between glutamine and glycine residues. This finding supports the hypothesis that the cleavage of VP0, which occurs during virion morphogenesis, is distinct from the cleavages that separate functional regions of the polyprotein.
McDonough feline sarcoma virus: characterization of the molecularly cloned provirus and its feline oncogene (v-fms)
Authors: Donner L , Fedele L A , Garon C F , Anderson S J , Sherr C J
Published: January 2020

The genetic structure of the McDonough strain of feline sarcoma virus (SM-FeSV) was deduced by analysis of molecularly cloned, transforming proviral DNA. The 8.2-kilobase pair SM-FeSV provirus is longer than those of other feline sarcoma viruses and contains a transforming gene (v-fms) flanked by sequences derived from feline leukemia virus. The order of genes with respect to viral RNA is 5'-gag-fms-env-3', in which the entire feline leukemia virus env gene and an almost complete gag sequence are represented. Transfection of NIH/3T3 cells with cloned SM-FeSV proviral DNA induced foci of morphologically transformed cells which expressed SM-FeSV gene products and contained rescuable sarcoma viral genomes. Cells transformed by viral infection or after transfection with cloned proviral DNA expressed the polyprotein (P170gag-fms) characteristic of the SM-FeSV strain. Two proteolytic cleavage products (P120fms and pp55gag) were also found in immunoprecipitates from metabolically labeled, transformed cells. An additional polypeptide, detected at comparatively low levels in SM-FeSV transformants, was indistinguishable in size and antigenicity from the envelope precursor (gPr85env) of feline leukemia virus. The complexity of the v-fms gene (3.1 +/- 0.3 kilobase pairs) is approximately twofold greater than the viral oncogene sequences (v-fes) of Snyder-Theilen and Gardner-Arnstein FeSV. By heteroduplex, restriction enzyme, and nucleic acid hybridization analyses, v-fms and v-fes sequences showed no detectable homology to one another. Radiolabeled DNA fragments representing portions of the two viral oncogenes hybridized to different EcoRI and HindIII fragments of normal cat cellular DNA. Cellular sequences related to v-fms (designated c-fms) were much more complex than c-fes and were distributed segmentally over more than 40 kilobase pairs in cat DNA. Comparative structural studies of the molecularly cloned proviruses of Synder-Theilen, Gardner-Arnstein, and SM-FeSV showed that a region of the feline-leukemia virus genome derived from the pol-env junction is represented adjacent to v-onc sequences in each FeSV strain and may have provided sequences preferred for recombination with cellular genes.
Nucleotide sequence of the 5' noncoding region and part of the gag gene of Rous sarcoma virus
Authors: Swanstrom R , Varmus H E , Bishop J M
Published: January 2020

Several functions of the retrovirus genome involve structural features in the vicinity of its 5' terminus. In an effort to further elucidate the relationship between structure and function in retrovirus RNA, we have determined the sequence of the first 1,010 nucleotides at the 5' end of the genome of Rous sarcoma virus by using the Maxam-Gilbert method to sequence suitable domains in cloned Rous sarcoma virus DNA. The results (i) locate the initiation codon for the gag gene of Rous sarcoma virus 372 nucleotides from the 5' end of viral RNA; (ii) demonstrate that this codon is preceded by three methionine codons that are apparently not used in translation; (iii) sustain previous conclusions that the principal site to which ribosomes bind on the Rous sarcoma virus genome in vitro does not contain the initiation codon for gag; (iv) permit deduction of the amino acid sequence of a viral structural protein, p19; (v) confirm the amino-terminal sequence of Pr76gag; and (vi) substantiate the identification of a splice donor site described in the accompanying manuscript (Hackett et al., J. Virol., 41:527-534, 1982).
Long terminal repeat of murine retroviral DNAs: sequence analysis, host-proviral junctions, and preintegration site
Authors: Van Beveren C , Rands E , Chattopadhyay S K , Lowy D R , Verma I M
Published: January 2020

The nucleotide sequence of the long terminal repeat (LTR) of three murine retroviral DNAs has been determined. The data indicate that the U5 region (sequences originating from the 5' end of the genome) of various LTRs is more conserved than the U3 region (sequences from the 3' end of the genome). The location and sequence of the control elements such as the 5' cap, "TATA-like" sequences, "CCAAT-box," and presumptive polyadenylic acid addition signal AATAAA in the various LTRs are nearly identical. Some murine retroviral DNAs contain a duplication of sequences within the LTR ranging in size from 58 to 100 base pairs. A variant of molecularly cloned Moloney murine sarcoma virus DNA in which one of the two LTRs integrated into the viral DNA was also analyzed. A 4-base-pair duplication was generated at the site of integration of LTR in the viral DNA. The host-viral junction of two molecularly cloned AKR-murine leukemia virus DNAs (clones 623 and 614) was determined. In the case of AKR-623 DNA, a 3- or 4-base-pair direct repeat of cellular sequences flanking the viral DNA was observed. However, AKR-614 DNA contained a 5-base-pair repeat of cellular sequences. The nucleotide sequence of the preintegration site of AKR-623 DNA revealed that the cellular sequences duplicated during integration are present only once. Finally, a striking homology between the sequences flanking the preintegration site and viral LTRs was observed.
SP01 gene 27 is required for viral late transcription
Authors: Greene J R , Chelm B K , Geiduschek E P
Published: January 2020

The SP01 mutant sus HA20 (gene 27) was found to be defective for synthesis of viral late RNA. It is known that gene 27 is also required for viral DNA replication. The SP01 gene 27 product resembles the T4 gene 45 product, which also has a dual role in viral DNA replication and late transcription.
MB78, a virulent bacteriophage of Salmonella typhimurium
Authors: Joshi A , Siddiqi J Z , Rao G R , Chakravorty M
Published: January 2020

The isolation and some properties of a virulent bacteriophage of Salmonella typhimurium, MB78, which is morphologically, serologically, and physiologically unrelated to P22, are reported. The phage has a noncontractile long tail with partite ends. It cannot multiply in minimal medium in the presence of citrate. MB78-infected cells are, however, killed in such medium. This phage cannot grow in rifampin-resistant mutants of the host. The latent period of growth of this phage is much shorter than that of P22. Both sieA and sieB genes of the resident P22 prophage are required to exclude the superinfecting MB78 phage, whereas all temperate phages related to P22 are excluded by either one or both of the genes individually. Restriction endonuclease cleavage patterns of P22 and MB78 are distinctly different. The absence of homology between the two phages P22 and MB78 suggests that MB78 is not related to phage P22.
Purification of glycoprotein gD of herpes simplex virus types 1 and 2 by use of monoclonal antibody
Authors: Eisenberg R J , Ponce de Leon M , Pereira L , Long D , Cohen G H
Published: January 2020

Glycoproteins gD-1 and gD-2 of herpes simplex virus types 1 and 2, respectively, were purified on an immunoadsorbent consisting of the type-common monoclonal antibody HD-1 linked to Sepharose. Each glycoprotein was of sufficient purity, quantity, and biological activity to be used for immunological and biochemical studies. Each glycoprotein induced high titers of type-common monospecific neutralizing antibody in mice. Amino aicd analysis indicated that gD-1 and gD-2 had similar though not identical amino acid compositions.
Intracisternal A-particle genes: structure of adjacent genes and mapping of the boundaries of the transcriptional unit
Authors: Cole M D , Ono M , Huang R C
Published: January 2020

Adjacent intracisternal A-particle (IAP) genes were identified in two different recombinant DNA clones, gamma 81 and gamma 19. In clone gamma 81, the most common form of IAP gene was separated by 5.3 kilobases from another IAP gene that had two apparent internal deletions. The two genes were in a head-to-tail configuration. In clone gamma 19, two different types of IAP genes were separated by less than 0.5 kilobase. Blot hybridization analysis of mouse DNA demonstrated that the DNA sequence found in clone gamma 81 is identical to the in vivo configuration. Using isolated DNA fragments from clone gamma 19, we mapped the boundaries of the IAP RNA by S1 digestion of RNA-DNA hybrids and by cDNA extension. With these techniques, both the 5' end and the 3' end of the IAP RNA in two different plasmacytomas (MOPC 315 and TEPC 15) were shown to fall within the long terminal direct repeat of the IAP gene. The fragment sizes generated by S1 digestion of IAP RNAs isolated from the two tumor lines were found to differ, indicating that different IAP genes may be transcribed in these two plasmacytomas.