bluuebunny/crossref_metadata_2025 huggingface dataset:
Evolutionary relationships of the primate papovaviruses: base sequence homology among the genomes of simian virus 40, stump-tailed macaque virus, and SA12 virus
Authors: Newell N , Shah K V , Kelly T J
Published: January 2020

Physical maps of the genomes of the two newly discovered primate papovaviruses, SA12 and stump-tailed macaque virus (STMV), were generated by restriction endonuclease analysis. The base sequence homologies among the genomes of SA12, stump-tailed macaque virus, and simian virus 40 (SV40) were studied by heteroduplex analysis. Heteroduplexes between SA12 and SV40 DNAs and stump-tailed macaque virus and SV40 DNAs were constructed and mounted for electron microscopy in various amounts of formamide to achieve a range of effective temperatures. At each effective temperature, the regions of duplex DNA in the heteroduplexes were measured and localized on the SV40 physical and functional maps. By analyzing the data from this study and rom our previous study (N. Newell, C. J. Lai, G. Khoury, and T. J. Kelly Jr., J. Virol. 25:193-201, 1978) on the base sequence homology between the genomes of BK virus and SV40, some general conclusions have been drawn concerning the evolutionary relationships among the genomes of the primate papovaviruses. The extent of homology among the viral genomes does not reflect the phylogenetic relationships of their hosts. At comparable effective temperatures Tm - 33 degrees C), the heteroduplexes between the DNAs of BK virus and SV40 contained the largest amount of duplex (about 90%). The heteroduplexes made between SA12 and SV40 DNAs were slightly less homologous, containing about 80% duplex. The heteroduplexes made between SV40 and stump-tailed macaque virus DNAs were only 20% duplex under the same conditions. When the various heteroduplexes were mounted for microscopy at effective temperatures greater than Tm - 33 degrees C, the fraction of the duplex DNA decreased in each case, indicating the existence of considerable base mismatching in the homologous regions. When specific coding or noncoding regions of the viral genomes were compared, the data indicated that the extent of sequence divergence differed markedly from one region to another. In all the heteroduplexes studied, there were two regions, located near the junctions between early and late regions on the SV40 map, which were essentially nonhomologous. All of the heteroduplexes studied showed significantly greater homology in the late region than in early region. Within the late region, the sequences coding for the major capsid polypeptide, VP1, were the most highly conserved.
Morphogenesis of foot-and-mouth disease virus. I. Role of procapsids as virion Precursors
Authors: Yafal A G , Palma E L
Published: January 2020

The role of procapsids during foot-and-mouth disease virus multiplication was studied on infected BHK-21 cells. Purified virus and procapsids were obtained by treating the infected cytoplasmic extracts with RNase and EDTA. The synthesis of virus, procapsids, and total particles was determined in pulse-chase experiments. A precursor-product relationship between procapsids and virions was obtained. The results show that the rate of synthesis of total particles (virus + procapsids) was linear from the addition of the label and was identical to that corresponding to virions. Therefore, the speed of the morphogenetic process as well as the existence of a precursor pool of structural proteins was established. Furthermore, the rate of virus synthesis from procapsids was identical to the rate of synthesis of procapsids from their structural precursors. A quantitative recovery of label from procapsids into virions was obtained by the use of cycloheximide or tosyl-lysine chloromethyl ketone. Under these conditions, virus synthesis proceeds, indicating that these drugs do not affect the morphogenetic step studied in this paper.
Simian Virus 40 Mutants with Deletions at the 3′ End of the Early Region Are Defective in Adenovirus Helper Function
Authors: Cole Charles N. , Crawford Lionel V. , Berg Paul
Published: January 2020

Coinfection of monkey cells with simian virus 40 (SV40) and adenovirus type 2 (Ad2) increased the Ad2 yield 1,000-fold over that obtained by Ad2 infection alone of monkey cells (A. S. Rabson, G. T. O'Conor, I. K. Berezesky, and F. J. Paul, Proc. Soc. Exp. Biol. Med.
116
:187-190, 1964). The ability of viable mutants of SV40 that contain deletions at various sites in the viral DNA to enhance Ad2 growth in monkey cells was examined. Only those mutants with deletions near the 3′ end of the early region were deficient in providing this helper function. Mutants
dl
1265, lacking 39 base pairs at map position 0.18, and
dl
1263, lacking 33 base pairs at map position 0.20 (H. van Heuverswyn, C. Cole, P. Berg, and W. Fiers, J. Virol.
30
:936-941, 1979), were approximately 4 and 30% as effective as wild-type SV40, respectively. The extent of enhancement of Ad2 yield depended on the multiplicity of infection by SV40, but not by Ad2 (at a multiplicity of infection of ≤50), as well as on the relative times of infection by Ad2 and SV40. Increasing the SV40 multiplicity of infection or infecting cells with SV40 wild type or mutants prior to Ad2 infection increased the Ad2 yield dramatically. The T antigens of wild-type SV40,
dl
1263, and
dl
1265 were examined. We attempt to correlate defects in helper function, alterations in the T antigen structure, and the DNA sequence of the mutants as determined by van Heuverswyn et al. (J. Virol.
30
:936-941, 1979).
Characterization of a protein found in cells infected with the spleen focus-forming virus that shares immunological cross-reactivity with the gp70 found in mink cell focus-inducing virus particles
Authors: Ruscetti S K , Linemeyer D , Feild J , Troxler D , Scolnick E M
Published: January 2020

Previously we detected an antigen in cells infected with the spleen focus-forming virus (SFFV) with a radioimmunoassay specific for the gp 70's of murine leukemia mink cell focus-inducing (MCF) viruses. This antigen has now been characterized in competition radioimmunoassays with limiting dilutions of antibody and in pulse-labeling studies under conditions of antibody excess. Both methods of analysis indicate that the SFFV-encoded antigen is a glycoprotein with a molecular weight of approximately 52,000. The gp52 shared immunological reactivity and methionine-containing tryptic peptides with the gp70 of a Friend MCF virus and was expressed on the surface of SFFV-infected cells as well as in the cytoplasm. The gp52 could be detected (i) in fibroblastic cell lines from several species when these cells were infected with SFFV; (ii) in several established erythroleukemic cell lines; and (iii) in the spleens of mice recently infected with SFFV. Although it shared immunochemical properties with the gp70 of Friend MCF virus, the gp52 could be distinguished from the MCF gp70 (i) by its apparent lack of group and interspecies immunological determinants compared with MCF virus-derived gp70's; (ii) by its failure to be released from cells infected with SFFV or SFFV plus helper virus; (iii) by its molecular weight; and (iv) by tryptic peptide analysis. The results indicate that SFFV codes for an MCF gp70-related gp52 which is apparently no longer a virion structural protein like the MCF gp70 from which it was originally derived.
Nature and distribution of feline sarcoma virus nucleotide sequences
Authors: Frankel A E , Gilbert J H , Porzig K J , Scolnick E M , Aaronson S A
Published: January 2020

The genomes of three independent isolates of feline sarcoma virus (FeSV) were compared by molecular hybridization techniques. Using complementary DNAs prepared from two strains, SM- and ST-FeSV, common complementary DNA'S were selected by sequential hybridization to FeSV and feline leukemia virus RNAs. These DNAs were shown to be highly related among the three independent sarcoma virus isolates. FeSV-specific complementary DNAs were prepared by selection for hybridization by the homologous FeSV RNA and against hybridization by fline leukemia virus RNA. Sarcoma virus-specific sequences of SM-FeSV were shown to differ from those of either ST- or GA-FeSV strains, whereas ST-FeSV-specific DNA shared extensive sequence homology with GA-FeSV. By molecular hybridization, each set of FeSV-specific sequences was demonstrated to be present in normal cat cellular DNA in approximately one copy per haploid genome and was conserved throughout Felidae. In contrast, FeSV-common sequences were present in multiple DNA copies and were found only in Mediterranean cats. The present results are consistent with the concept that each FeSV strain has arisen by a mechanism involving recombination between feline leukemia virus and cat cellular DNA sequences, the latter represented within the cat genome in a manner analogous to that of a cellular gene.
Glycopeptides of Murine Leukemia Viruses I. Comparison of Two Ecotropic Viruses
Authors: Kemp Maurice C. , Basak Sukla , Compans Richard W.
Published: January 2020

The glycopeptides obtained by pronase digestion of two ecotropic strains of murine leukemia virus (MuLV) were compared by gel filtration. Four different glycopeptide size classes, designated G
1
, G
2
, G
3
, and G
4
, with molecular weights of approximately 5,100, 2,900, 2,200, and 1,500, respectively, were shown to be associated with Rauscher MuLV virions grown in JLS-V9 cells. Various sugar precursors, including glucosamine, galactose, fucose, and mannose were incorporated into G
1
and G
2
, suggesting that these are complex (type I) glycopeptides. The two smaller glycopeptide size classes, G
3
and G
4
, were shown to be mannoserich (type II) glycopeptides. G
4
was more sensitive to digestion with endo-β-
N
-acetylglucosaminidase H than G
3
, suggesting that the core of G
3
may contain fewer mannose residues. Glycopeptides of the same size class as G
1
and G
2
were associated with both Rauscher MuLV and AKR-MuLV grown in III6A (mouse embryo) cells. Previous studies have shown that gp52, a proteolytic cleavage product of gp70, possessed primarily G
1
glycopeptides and that gp52 was more highly sulfated than gp70. We observed that G
1
is approximately twofold more highly sulfated than G
2
, explaining the observed difference in sulfation of gp52. The unusually large size of G
1
suggested that infection with MuLV may alter the host cell glycosylation pattern. To test this possibility, glycopeptides from Sindbis virions grown in uninfected and Rauscher MuLV-infected JLS-V9 cells were compared, and no differences were observed. G
1
was not detected in Sindbis virions, indicating that acquisition of G
1
depends on properties of the virus-coded polypeptide backbone of the gp70 molecule.
Biochemical characterization of temperature-sensitive rabies virus mutants
Authors: Saghi N , Flamand A
Published: January 2020

Biochemical characterization of 70 temperature-sensitive (ts) mutants of rabies virus has been done by following the appearance of viral proteins and RNA molecules in infected cells at both permissive and nonpermissive temperature. The presence or absence of the nucleocapsid protein (N) was demonstrated by treating infected cells with anti-N fluorescent antibodies. At 33 degrees C, all the mutants induced a fluorescence comparable to the wild type. At 39.6 degrees C, the mutants can be classified into three groups. Three mutants induced a fluorescence comparable to the wild type (F+ mutants); 54 mutants induced a faint fluorescence which was proportional to the multiplicity of infection and increased with time (F+- mutants). No fluorescence could be detected for the 13 remaining mutants (F- mutants). The synthesis of all viral proteins was shown to be normal for F+ mutants, indicating that transcription and replication of the virus were normal and that the ts lesion was located in a protein which is not directly required for those functions. The synthesis of all viral proteins was similarly decreased for F+- mutants and undetectable for the F- mutants. This suggests that the ts lesion affects the transcription and/or replication of the virus. By annealing techniques it was demonstrated that the F+- mutants were able to perform some amount of secondary transcription at nonpermissive temperature. No secondary transcription occurred with F- mutants. When detectable (i.e., at higher multiplicity of infection), primary transcription of F- mutants was normal.
Isolation, Complementation, and Initial Characterization of Temperature-Sensitive Mutants of the Baculovirus
Autographa californica
Nuclear Polyhedrosis Virus
Authors: Lee Hyung H. , Miller Lois K.
Published: January 2020

Sixteen temperature-sensitive mutants of
Autographa californica
nuclear polyhedrosis virus were isolated. Several interesting phenotypes were observed. A large proportion of the mutants were unable to form polyhedral occlusion bodies (polyhedra) at the nonpermissive temperature (32.5°C). At 32.5°C, one mutant formed plaques in which the cells lacked polyhedra. Another mutant type was defective in the production of progeny extracellular nonoccluded virus and produced a “plaque” consisting of only a single cell containing polyhedra at 32.5°C. One mutant was defective in plaque formation, progeny nonoccluded virus formation, and polyhedra formation at 32.5°C. Several mutants produced nonoccluded virus but failed to produce plaques or polyhedra at 32.5°C. Other phenotypes were also distinguished. Complementation analyses, performed by either measuring the increase in extracellular nonoccluded virus formation or by observing polyhedra formation in mixed infections at 32.5°C, indicated the presence of 15 complementation groups. A high frequency of recombination was observed. Four of the mutants were found to be host dependent in their temperature sensitivity for polyhedra formation.
Carbohydrates of influenza virus. III. Nature of oligosaccharide-protein linkage in viral glycoproteins
Authors: Keil W , Klenk H D , Schwarz R T
Published: January 2020

Four different glycopeptides can be distinguished after pronase digestion of influenza A virus glycoproteins: Ia and Ib, containing N-acetylglucosamine, mannose, galactose, and fucose, and IIa and IIb, containing mannose and N-acetylglucosamine. All glycopeptides yielded N-acetylglucosaminyl-asparagine after mild acid hydrolysis. There was no evidence for O-glycosidic bonds. Thus, the carbohydrate complement is linked to the polypeptide exclusively by N-glycosidic linkages between N-acetylglucosamine and asparagine.
High-frequency recombination within the gag gene of Rous sarcoma virus
Authors: Linial M , Brown S
Published: January 2020

We isolated 28 recombinants of Rous sarcoma virus at early (24 h) and late (7 days) times after infection. These recombinants were selected for wild type in the pol and src genes and analyzed for their env and gag phenotypes. We were unable to show strong linkage between any two markers, including two markers within a single gene (gag).
Inhibition of influenza virus uncoating by rimantadine hydrochloride
Authors: Koff W C , Knight V
Published: January 2020

In freeze-thaw lysates of MDCK cells infected with 32P-labeled influenza virus A/WSN in the presence of added RNase, acid-precipitable radioactivity diminished to about 50% of initial values within 90 min after a 1-h virus adsorption period. A similar preparation containing rimantadine at a concentration of 50 micrograms/ml exhibited only a 10% reduction in acid-precipitable radioactivity. These findings suggest that rimantadine interferes with uncoating of influenza virus in infected cells.
Diversity of mammary tumor viral genes within the genus Mus, the species Mus musculus, and the strain C3H
Authors: Drohan W , Schlom J
Published: January 2020

Proviral sequences complementary to the C3H mouse mammary tumor virus RNA genome are present in the DNA of early occurring mammary tumors of C3H/HeN mice and are absent from apparently normal C3H/HeN tissues; these sequences are non-germ line transmitted in C3H/HeN mice and have been termed tumor-associated sequences; (W. Drohan et al., J. Virol. 21:986-995, 1977). We report here that tumor-associated sequences are present in the DNA of spontaneous mammary tumors that occur early in the life of several inbred, high-tumor-incidence mouse strains but are absent in mammary tumors that occur later in life in low- and moderate-tumor-incidence strains. These sequences are also absent in apparently normal organs tested from numerous laboratory mouse strains, feral mice, Mus musculus subspecies, and other Mus species. Sequences represented in tumor-associated sequence RNA, however, are present as endogenous provirus in GR mice (at approximately four copies per haploid genome) and in two of five substrains of C3H mice tested (at approximately one copy per haploid genome). The two substrains of C3H mice positive for endogenous tumor-associated sequence provirus were recently (circa 1930) separated from the negative substrains of C3H mice. The results may be explained by the unlikely chance segregation of proviral sequences or by the recent integration of viral genes (within the last few decades). Whereas radioactively labeled mouse mammary tumor virus 60-70S RNA or complementary DNA detected mouse mammary tumor virus-related proviral information in all laboratory mouse strains, feral mice, subspecies of M. musculus, and other species of Mus, the use of tumor-associated sequence RNA clearly revealed the genetic diversity that may exist between different colonies or substrains of "inbred" laboratory mice commonly used in cancer research.
Physical mapping of paar mutations of herpes simplex virus type 1 and type 2 by intertypic marker rescue
Authors: Chartrand P , Stow N D , Timbury M C , Wilkie N M
Published: January 2020

Mutations (paar) in herpes simplex virus (HSV) which confer resistance to phosphonoacetic acid involve genes associated with virus-induced DNA polymerase activity. Two mutants of HSV (HSV-1 tsH and HSV-2 ts6) produce a thermolabile DNA polymerase activity. In this study, the ts lesions present in these mutants and those present in two independent phosphonoacetic acid-resistant mutants of HSV-1 and HSV-2 (paar-1 and paar-2) have been physically mapped by restriction endonuclease analysis of recombinants produced between HSV-1 and HSV-2 by intertypic marker rescue. All four mutations mapped within a 3.3-kilobase pair region around map unit 40. The accuracy of the method is reflected by the mapping results for tsH and paar-2, which were found to lie in the same 1.3-kilobase pair region. paar-1 was found to lie to the right of ts6. Virus-induced DNA polymerase is thought to have a molecular weight of 150,000, necessitating a gene with a coding capacity of 4.6 kilobase pairs. The four mutations mapped in this study all lie within a region smaller than this, but the results do not yet prove that all four lesions reside in this or any single gene.
DNA of Epstein-Barr virus. VI. Mapping of the internal tandem reiteration
Authors: Given D , Kieff E
Published: January 2020

Epstein-Barr virus (B95-8) DNA consists of short (10 X 10(6)) and long (87 X 10(6)) unique DNA sequences joined by 10 tandem reiterations of a 1.85 X 10(6) DNA segment. The reiterated sequence contains BamI and BglII sites separated by 4 X 10(5). The 4.5 X 10(5) and 14.0 X 10(5) segments generated by cleavage of the reiterated DNA with BamI and BglII contain sequences which hybridize to each other, suggesting that the internal tandemly reiterated sequence has a direct or inverted repeat within it. The opposite ends of the linear, nicked, double-stranded DNA molecule (R. F. Pritchett, S. D. Hayward, and E. D. Kieff, J. Virol. 15:556--569, 1975) consist of from 1 to 12 direct repeats of another 3 X 10(5) sequence (D. Given and E. Kieff, J. Virol. 28:524--542, 1978; D. Given, D. Yee, K. Griem, and E. Kieff, J. Virol. 30:852--862, 1979). There is no homology between the internal reiterated sequence and either terminus. However, part of the internal reiteration (less than 5 X 10(5) is reiterated at two separate locations in the long unique region. The internal reiterations are a source of variation within EBV (B95-8) DNA preparations. Thus, although the majority of molecules contain 10 tandem reiterations, some molecules have 9, 8, 7, 6, 5, 4, or fewer tandem reiterations. A consequence of this variability is that the KpnI A fragment and the EcoRI/Hsul A fragment consist of a family of seven or more fragments differing in the number of tandem internal reiterations. The EcoRI/HsuI A fragment of EBV (W91) DNA is approximately 6 X 10(6) smaller than the largest and dominant EcoRI/HsuI A fragment of EBV (B95-8) DNA. EBV (W91 DNA also differs from EBV (B95-8) DNA by an additional 7 X 10(6) to 8 X 10(6) of DNA in the long unique DNA region (D. Given and E. Kieff, J. Virol. 28:524--542, 1978; N. Raab-Traub, R. Pritchett, and E. Kieff, J. Virol. 27:388--398, 1978). These data suggest the possibility that the smaller number of internal reiterations in EBV (W91) DNA may be a consequence of the additional unique DNA and a restriction in the overall size of EBV DNA.
Temperature-Sensitive Mutants of Complementation Group E of Vesicular Stomatitis Virus New Jersey Serotype Possess Altered NS Polypeptides
Authors: Evans D. , Pringle C. R. , Szilágyi J. F.
Published: January 2020

In vesicular stomatitis virus New Jersey serotype polyacrylamide gel electrophoresis was unable to distinguish the polypeptides of the temperature-sensitive (
ts
) mutants of complementation groups A, B, C, and F from those of the wild-type virus. However, the NS polypeptide of the representative mutant of group E,
ts
E1, had a significantly greater electrophoretic mobility than that of the wild-type virus NS polypeptide. The electrophoretic mobilities of the NS polypeptides of the three mutants of complementation group E varied, being greatest in the case of
ts
E1, slightly less for
ts
E2, and only a little greater than that of wild-type virus NS polypeptide in the case of
ts
E3. Since the NS polypeptides of the revertant clones
ts
E1/R1 and
ts
E3/R1 have mobilities identical to that of wild-type NS polypeptide, the observed altered mobilities of the group E mutants are almost certainly the direct result of the
ts
mutations in the E locus. The electrophoretic mobilities of the intracellular NS polypeptides of the group E mutants were indistinguishable from those of their virion NS polypeptides. The electrophoretic mobilities of the NS polypeptides of the group E mutants synthesized in vitro using mRNA synthesized in vitro by TNP were identical to those of the NS polypeptides of their purified virions. The NS polypeptides of all three mutants were labeled with
32
P
i
to approximately the same extent as wild-type virus NS polypeptide, indicating that gross differences in phosphorylation of this polypeptide are unlikely to account for the altered mobilities. We propose a model in which the NS polypeptide consists of at least three loops held in this configuration by hydrophobic or ionic forces or both and stabilized by phosphodiester bridges. If a mutation affects one of the amino acids to which the phosphate is covalently linked, the phosphodiester bridge cannot be formed, and, as a result, in the presence of sodium dodecyl sulfate the affected loop opens and thus the NS polypeptide migrates further into the gel. Such a configuration may also explain the multifunctional nature of the NS polypeptide.
Radioimmunoassays for the 36,000-dalton glycoprotein of murine mammary tumor viruses demonstrate type, group, and interspecies determinants
Authors: Teramoto Y A , Schlom J
Published: January 2020

Mouse mammary tumor viruses (MMTVs) contain distinct membrane glycoproteins of 52,000 daltons (gp52) and 36,000 daltons (gp36). We report here the development of new radioimmunoassays for gp36, using gp36 purified by hydrophobic chromatography and gel filtration. These assays demonstrate that gp36 has both type-specific and group-specific antigenic determinants. The virus-coded nature of these determinants was shown by utilizing different MMTVs grown in the same feline cell line. Interspecies determinants on gp36 were demonstrated by the observations that (i) MC-MTV (a virus isolate from the Asian rodent Mus cervicolor, and morphologically identical to MMTVs) competed, with an altered slope, in the gp36 radioimmunoassay, and (ii) antisera raised against MC-MTV immunopreciptitated 125I-labeled gp36. The detection of gp36 in spontaneous mammary tumors of several strains of mice also facilitates further studies on the replication of MMTVs and the host's immune response to MMTV-mediated oncogenesis.
Simultaneous Purification of Murine Mammary Tumor Virus Structural Proteins: Analysis of Antigenic Reactivities of Native gp34 by Radioimmunocompetition Assays
Authors: Marcus Stuart L. , Kopelman Rebecca , Sarkar Nurul H.
Published: January 2020

All the structural proteins (gp47, gp34, p27, p23, p16, and p12) of the murine mammary tumor virus (MuMTV) were simultaneously purified utilizing alkylagarose chromatography as the initial fractionation step. Least-hydrophobic MuMTV polypeptides (p23, p16) and the slightly hydrophobic p27 were separated from moderately hydrophobic proteins gp47 and p12 by passage through octylimino (C
8
)-agarose; the gp47 and p12 could be removed from the matrix by elution with ethylene glycol, whereas the most hydrophobic MuMTV protein, gp34, was eluted using nonionic detergent together with ethylene glycol. Subsequent purification steps involved ion-exchange or gel filtration chromatography. The resulting protein preparations appeared near-homogeneous on analysis by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Recoveries of MuMTV proteins, based on their approximate individual contribution to total virus protein, ranged from about 20% for gp47 to greater than 100% for the minor structural component p23, the major phosphoprotein of MuMTV. Antiserum against purified C3H MuMTV gp34, together with purified, radioiodinated gp34, was used to develop a radioimmunoassay which showed that from 13 to 14% of total MuMTV protein by weight is gp34. Using this assay system, the group-specific antigenic reactivity of gp34 was also demonstrated. When solubilized preparations of C3H, RIII, and GR MuMTV's were used as competing antigens in gp34 radioimmunoassays with anti-C3H MuMTV serum, both group- and type-specific differences in antigenic reactivity were found.
RNA-directed DNA synthesis in Moloney murine leukemia virus: interaction between the primer tRNA and the genome RNA
Authors: Peters G , Dahlberg J E
Published: January 2020

Initiation of RNA-directed DNA synthesis in virions of Moloney murine leukemia virus requires a cellular tRNAPro as primer. The site(s) on the Moloney murine leukemia virus genome RNA at which functional primer molecules are bound and at which purified tRNAPro hybridizes has been located near (within 20%) the 5' end of the genome. A relatively stable duplex (temperature at which 50% dissociation has occurred, 76 degrees C) is formed between the amino acid acceptor stem of the tRNAPro and a complementary sequence in the Moloney murine leukemia virus 35S RNA. The interaction involves 19 base pairs, extending from the penultimate nucleotide at the 3' end of the tRNAPro but apparently not including the 3'-terminal adenosine residue. In most respects, the interaction between primer and template in Moloney murine leukemia virus parallels the situation in the avian leukosis-sarcoma viruses.
Specific neutralization of defective spleen focus-forming virus in Friend virus complex by rat antiserum
Authors: Yoosook C , Steeves R A , Ruscetti S
Published: January 2020

The spleen focus-forming virus (SFFV), a rapidly transforming, replication-defective virus in Friend virus (FV) complex that is readily neutralized by antisera directed against its helper virus, was examined for the presence of SFFV-specific antigens. Antisera prepared in Fisher rats against an SFFV-infected Fisher rat embryo fibroblast line (SFFV-FRE) neutralized SFFV effectively, but not Friend-associated murine leukemia virus (F-MuLV) whether the latter was tested alone or was mixed with SFFV in the FV complex. In contrast, serum from mice immunized with SFFV-infected nonproducer mouse cells had little or no neutralizing activity against SFFV. Both absorption and immunoprecipitation studies indicate that the SFFV-specific antigen is immunologically related to xenotropic murine leukemia virus antigens. The role of both SFFV- and F-MuLV-specific antigens in the neutralization of SFFV suggests that this defective virus could be an antigenic mosaic and that viruses in the FV complex may participate in a undirectional form of phenotypic mixing.
Further characterization of the replicative complex of vesicular stomatitis virus
Authors: Simonsen C C , Hill V M , Summers D F
Published: January 2020

Replicating vesicular stomatitis virus ribonucleoprotein (RNP) complexes were isolated in nonequilibrium Renografin density gradients. These nascent RNPs had the same buoyant density as virion nucleocapsids in both isopycnic Renografin and CsCl gradients. Both transcribing and replicating RNP complexes were shown to be stable in sucrose gradients, whereas only replicating RNP complexes were stable in Renografin gradients. Size analysis of the 5-min-pulse-labeled RNA species from the replicating RNPs using methylmercury gels revealed that the nascent strands were primarily less than full-length molecules. Longer times of radiolabeling demonstrated that the nascent RNA accumulated as 42S RNA, which was primarily of the same sense as the virion strand when it was radiolabeled at 5 h postinfection. The percentage of this radiolabeled RNA which was plus stranded was higher at 2.5 h postinfection, reflective of the shift in plus- to minus-stranded full-length 42S RNA synthesis which occurs in the cell. Addition of cycloheximide to the infected cells before the addition of the radiolabel prevented the formation of these RNP complexes. Both the change in the percentage of minus strands found in the RNP complexes at the different times postinfection and the sensitivity to cycloheximide indicate that the RNP complex which was isolated was indeed the replicative complex.
p21 of Kirsten murine sarcoma virus is thermolabile in a viral mutant temperature sensitive for the maintenance of transformation
Authors: Shih T Y , Weeks M O , Young H A , Scolnick E M
Published: January 2020

We have recently described an intracellular protein, p21, in nonproducer cells transformed by either the Kirsten (Ki-MSV) or Harvey (Ha-MSV) strain of murine sarcoma virus (Shih et al., Virology, in press). The p21 is phosphorylated and has been shown to be coded for by either Ki-MSV or Ha-MSV. In this report, we compare the thermal stability of the newly synthesized [35S]methionine-labeled p21 in cells transformed by the wild-type Ki-MSV or by a mutant of Ki-MSV (ts 371) which is temperature sensitive in a viral function required for the maintenance of several properties of the transformed phenotype. The immunoprecipitability of the p21 coded for by the ts 371 Ki-MSV was markedly more thermolabile than the p21 of the wild-type Ki-MSV when the cell extracts are heated in vitro. The present finding suggests that the p21 is required for the maintenance of transformation induced by Ki-MSV.
Virus production by Abelson murine leukemia virus-transformed lymphoid cells
Authors: Shields A , Rosenberg N , Baltimore D
Published: January 2020

Cell lines obtained by in vitro transformation of bone marrow with Abelson murine leukemia virus (A-MuLV) can be divided into three classes: producers, releasing reverse transcriptase-containing particles and infectious virus; nonproducers, releasing no viral particles; and defective producers, the most common phenotype, releasing particulate reverse transcriptase in the absence of infectious virus. When such cell lines were analyzed 1 to 2 weeks after their isolation, however, all produced infectious virus. Because these cell lines were carried in culture, many ceased to release infectious virus but produced defective virions. One defective producer, SWR4, has been extensively studied. The particles it produces have the same density as that of virions of Moloney murine leukemia virus (M-MuLV). The particles contain no 35 to 70S RNA, as determined by analysis of [3H]uridine-labeled particles, and exhibit no endogenous reverse transcriptase activity. Although the reverse transcriptase enzyme is of normal size, the major structural protein of the defective virions has a molecular weight of 28,000 (p28), in contrast to the p30 of M-MuLV, and no viral glycoprotein was evident. The defective particles do not appear to arise either from the helper virus or from Abelson virus. An alteration of the protein of the helper virus is an unlikely source of p28 because particles produced by lymphoid cells transformed with another strain of M-MuLV as helper (M-MuLV-TB) contained p28 with an unaltered cleavage pattern, although M-MuLV-TB p30 differs from M-MuLV p30. The A-MuLV genome lacks the capacity to code for the reverse transcriptase virions. Clones of fibroblasts infected with A-MuLV only occasionally produce defective particles. The defective particles therefore probably arose from an endogenous virus that is preferentially expressed in the class of lymphoid cells transformed by A-MuLV. This interpretation implies that the majority of A-MuLV-transformed lymphoid cells completely lose expression of the helper virus genome.
The genome of infectious bursal disease virus consists of two segments of double-stranded RNA
Authors: Müller H , Scholtissek C , Becht H
Published: January 2020

The RNA of infectious bursal disease virus was reexamined in a detailed analysis. It could be established that its genome consists of two segments of double-stranded RNA. The RNA is RNase resistant and has a sedimentation coefficient of 14S and a buoyant density of 1.62 g/ml. The purine/pyrimidine ratio is nearly 1; the guanine plus cytosine content is 55.3%; the Tm is 95.5 degrees C. The molecular weights of the two double-stranded segments were determined to be 2.2 x 10(6) and 2.5 x 10(6).
In vitro translation products specified by the transforming region of adenovirus type 2
Authors: Halbert D N , Spector D J , Raskas H J
Published: January 2020

Region 1 DNA sequences (map positions 0 to 11% on the linear adenovirus 2 genome) are expressed both early and late in lytic infection and are required for transformation by the virus. During productive infection six distinct cytoplasmic RNAs are synthesized from this region. These RNAs comprise two families, each consisting of three size classes that share 3' sequences. Region 1 RNA's were purified by hybridization selection, using restriction fragments bound to nitrocellulose membranes, and by size fractionation. The isolated RNAs were then translated in cell-free systems derived from wheat germ and rabbit reticulocytes. The family of RNAs specified by 0 to 4.4 sequences includes two RNAs, which are 12S and 13S in size. These RNAs were partially separated by molecular weight and translated. The 13S RNA produced 53,000-dalton (53K) and 41K peptides, and the 12S RNA synthesized 47K and 35K products. The family of RNAs mapping from 4.4 to 11.0 encodes three separate polypeptides, each of which can be assigned to a specific RNA. A 12K product that comigrates with structural polypeptide IX is synthesized from the 9S RNA as previously reported (U. Pettersson and M. B. Mathews, Cell 12:741-750, 1977). The 13S RNA encodes a 15K polypeptide that corresponds to a 15K polypeptide in infected cell extracts. The 22s RNA encodes a 52K protein distinct from the 0 to 4.4 polypeptides.
Effect of viral RNase H on the avian sarcoma viral genome during early transcription in vitro
Authors: Friedrich R , Moelling K
Published: January 2020

We investigated the influence of viral RNase H on the transcription of the avian sarcoma virus RNA in a virion-associated reaction. The ability of RNase H to degrade the RNA moiety of the initially formed RNA-DNA hybrid at the 5' end of the viral genome was found to be greatly dependent on the exact concentration of nonionic detergent used to activate the reaction. At a detergent concentration optimal for extensive and faithful in vitro transcription of avian sarcoma virus RNA by the virion-associated RNA-dependent DNA polymerase, most of the 5' terminus of the RNA was digested in 30 min at 41 degrees C. At higher than optimal detergent concentrations, however, little of that RNA was digested. We conclude that removal of the 5'-terminal redundancy in the RNA after its transcription into DNA is a prerequisite for base pairing of the DNA to the 3'-terminal redundant sequence. Lack of removal of this sequence leads to incorrect elongation and substantial reduction of DNA synthesis. When tested with a synthetic RNA-DNA hybrid, virion-associated RNase H did not reveal a detergent dependence.
Measles Virus-Specified Polypeptide Synthesis in Two Persistently Infected HeLa Cell Lines
Authors: Wechsler Steven L. , Rustigian Robert , Stallcup Kathryn C. , Byers Karen B. , Winston Stuart H. , Fields Bernard N.
Published: January 2020

Measles virus-directed protein synthesis was examined in two HeLa cell lines (K11 and K11A) that are persistently infected with wild-type measles virus. Four viral proteins (H, hemagglutination protein; P, nucleocapsid-associated protein; NP, the major nucleocapsid protein; and M, the matrix protein) were readily detected in both cell lines by immune precipitation of [
35
S]methionine-labeled cell extracts followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, three (H, NP, and M) of the four viral proteins in both K11 and K11A cells differed from the corresponding viral proteins synthesized in HeLa cells acutely infected with the parental wild-type virus. In addition, the M protein from K11A cells migrated significantly more slowly on sodium dodecyl sulfate-polyacrylamide gel electrophoresis than the M protein from K11 cells, and there appeared to be slight differences in the H and NP proteins between these two persistently infected cell lines. The altered viral proteins detected in K11 and K11A cells appeared to be the result of viral mutations rather than changes in the host cell, since virus recovered from these cells directed the synthesis of similar aberrant viral proteins in HeLa cells. Virus recovered from K11 cells and virus recovered from K11A cells were both temperature sensitive and grew more slowly than wild-type virus. HeLa cells infected with virus recovered from K11 cells readily became persistently infected, resembling the original persistently infected K11 cells. Thus, viral mutations are associated with persistent measles virus infections in cell cultures.
Genome complexities of the three mRNA species of snowshoe hare bunyavirus and in vitro translation of S mRNA to viral N polypeptide
Authors: Cash P , Vezza A C , Gentsch J R , Bishop D H
Published: January 2020

The genome complexities of the principal intracellular viral complementary RNA species of the snowshoe hare bunyavirus have been analyzed by duplex analyses involving hybridization of complementary RNA to individual 32P-labeled viral RNA species (large, L; medium, M; and small, S), recovery of nuclease-resistant duplexes, and determination of the oligonucleotide fingerprints of the protected 32P-labeled viral sequences. The result for the M RNA (which codes for the glycoproteins G1 and G2; J. R. Gentsch and D. H. L. Bishop, J. Virol. 30:767-770, 1979) indicates that there is a single polycistronic M mRNA. Similar results were obtained for the L and S RNA species. In vitro translation studies with the S complementary RNA species of snowshoe hare virus as well as melted purified S duplexes substantiate earlier genetic and molecular studies (J. R. Gentsch and D. H. L. Bishop, J. Virol. 28:417-419, 1978; J. Gentsch, D. H. L. Bishop, and J. F. Obijeski, J. Gen. Virol. 34-257-268, 1977), which indicate that S mRNA codes for the virion nucleocapsid protein N.
Recombination between snowhoe hare and La Crosse bunyaviruses
Authors: Gentsch J R , Robeson G , Bishop D H
Published: January 2020

We have previously reported heterologous genetic recombination resulting from crosses involving temperature-sensitive (ts) mutants of La Crosse (LAC) group II and snowshoe hare (SSH) group I ts mutants (J. Gentsch, L. R. Wynne, J. P. Clewley, R. E. Shope, and D. H. L. Bishop, J. Virol. 24:893-902, 1977). From those crosses two reassortant viruses having the large/medium/small viral RNA segment genotypes of SSH/LAC/SSH and SSH/LAC/LAC were obtained. In this study it has been found that the reciprocal cross (SSH group II x LAC group I ts mutants) has not yielded the expected LAC/SSH/SSH or LAC/SSH/LAC reassortant viruses. The backcross of a SSH/LAC/SSH group II ts mutant with a LAC group I ts mutant has produced a new reassortant virus, LAC/LAC/SSH, whereas the backcross of SSH/LAC/LAC group I ts mutants with SSH group II ts mutants gave another reassortant, SSH/SSH/LAC. Backcross analyses of LAC/LAC/SSH group I ts mutants with Group II ts mutants of SSH have not yielded the expected LAC/SSH/SSH reassortant virus, nor have backcrosses of SSH/SSH/LAC group II ts mutants with group I ts mutants of LAC virus yielded the expected LAC/SSH/LAC reassortant. Possible reasons why certain reassortant viruses are not produced are discussed. A procedure to screen SSH-LAC reassortant viruses which differ in their virion N polypeptides is described.
Heteroduplex Analysis of the Sequence Relationships Between the Genomes of Kirsten and Harvey Sarcoma Viruses, Their Respective Parental Murine Leukemia Viruses, and the Rat Endogenous 30S RNA
Authors: Chien Yueh-Hsiu , Lai Michael , Shih Thomas Y. , Verma Inder M. , Scolnick Edward M. , Roy-Burman Pradip , Davidson Norman
Published: January 2020

The sequence relations between Kirsten murine sarcoma virus (Ki-SV), Harvey murine sarcoma virus (Ha-SV), and a rat endogenous 30S RNA were studied by electron microscope heteroduplex analysis. The sequence relationships between the sarcoma viruses and their respective parental murine leukemia viruses (Kirsten and Moloney murine leukemia viruses), as well as between the two murine leukemia viruses, were also studied. The only observed nonhomology feature of the Kirsten murine leukemia virus/Moloney murine leukemia virus heteroduplexes was a substitution loop with two arms of equal length extending from 1.80 ± 0.18 kilobases (kb) to 2.65 ± 0.27 kb from the 3′ end of the RNA. It is believed that this feature lies in the
env
gene region of the viral genomes. The Ha-SV and Moloney murine leukemia virus genomes (respective lengths, 6.0 and 9.0 kb) were homologous in a 1.0 ± 0.05-kb region at the 3′ end and possibly over a 200-nucleotide region at the 5′ ends; otherwise, they were nonhomologous. Ha-SV and Ki-SV (length, 7.5 kb) were homologous in the first 4.36 ± 0.37-kb region from the 3′ end and in a 0.70 ± 0.15-kb region at the 5′ end. In between, there was a nonhomology region, possibly containing a short (0.23-kb) region of partial or total homology. The heteroduplex analysis between rat endogenous 30S RNA and Ki-SV shows that there are mixed regions of sequence homology and nonhomology at both the 5′ and 3′ ends. However, there is a large (4-kb) region of homology between Ki-SV and the rat 30S RNA in the center of the genomes, with only a small nonhomology hairpin feature. These studies help to define the regions of homology between the Ha-SV and Ki-SV genomes with each other and with the rat endogenous 30S RNA. These regions may be related to the sarcoma genicity of the viruses. In particular, the 0.7-kb region of homology of Ha-SV with Ki-SV at the 5′ ends may be related to the formation of a 21,000-dalton phosphoprotein in cells transformed by either virus.
Clonal analysis of mammalian cell cultures persistently infected with Japanese encephalitis virus
Authors: Schmaljohn C S , Blair C D
Published: January 2020

More than 200 cells were cloned from populations of mammalian cells persistently infected with Japanese encephalitis virus. Only four cloned cultures contained cells that had viral antigen measurable by immunofluorescence and that released infectious virus, yet all clones harbored virus-specific RNA. Superinfection of cloned cells with wild-type Japanese encephalitis virus did not produce cytopathic effects, but resulted in production of viral antigen and infectious virus in formerly nonproducing clones. Cocultivation of nonproducer clone cells with normally permissive cells did not induce virus production, nor did treatment of nonproducer clones with various inhibitors of DNA, RNA, or protein synthesis. It is suggested that the cloning procedure may have selected for a particular subpopulation of cells and that defective virus is also involved in establishment and maintenance of persistent infection.
The 55K protein on the 5' termini of adenovirus type 2 DNA is unrelated to virus-coded candidate transformation proteins (E1-53K, E1-40K-50K) and DNA-binding proteins (E2-42K/47K/73K)
Authors: Green M , Wold W S , Brackmann K H , Cartas M A
Published: January 2020

A polypeptide of 55,000 daltons (55K) is linked, probably covalently, to the K' termini of adenovirus type 2 DNA. The 55K polypeptide is synthesized during early stages of infection (T. Yamashita, M. Arens, and M. Green, J. Virol. 30: 497-507, 1979) and thus may function in viral DNA replication, gene regulation, or cell transformation. Several virus-coded early polypeptides have been identified that could correspond to the terminal 55K, including the E1-40K-50K and E1-53K candidate transformation polypeptides and the E2-42K/47K/73K single-stranded DNA-binding polypeptide. We show here that two-dimensional tryptic [35S]methionine-peptide maps of the terminal 55K differ completely from [35S]methionine-peptide maps of four related E1-40K-50K polypeptides, the E1-53K, and the related E2-42K, E2-47K, and E2-73K polypeptides. We conclude that the terminal 55K polypeptide does not correspond to any of the known virus-coded early polypeptides.
Characterization of infectious oncornaviruses from MOPC-460 plasmacytomas: their relation to A-type particles
Authors: Ramabhadran T V , Hartley J W , Rowe W P , Godefroy-Colburn T , Jhabvala P S , Thach R E
Published: January 2020

MOPC-460 mouse plasmacytoma cells produce intracellular A-type particles and extracellular oncornavirus-like particles ("myeloma-associated virus," abbreviated MAV). The genomes of these two particles are closely related. During attempts to establish infections with MOPC-460 extracellular particles, we isolated ecotropic and xenotropic infectious forms of murine leukemia virus. We have investigated the relation of these isolates to A-type particles and to MAV by nucleic acid hybridization. Using complementary DNA probes prepared from the two isolates, we found that these infectious murine leukemia viruses differ from A-type particles and from MAV. Moreover, we found that MAV is the predominant extracellular component: the ecotropic and xenotropic forms of murine leukemia virus were present at only low levels (less than 5%) in MAV preparations. Neither the SC-1 cells infected with ectropic murine leukemia virus nor the mink cells infected with xenotropic murine leukemia virus showed any A-type particles in their cytoplasm when examined by electron microscopy. Our inability to demonstrate infection by the A-type particle-related component, MAV, suggests that these may be defective.