bluuebunny/crossref_metadata_2025 huggingface dataset:
Avian myeloblastosis provirus cloned in a lambda bacteriophage is leukemogenic
Authors: Silva R F , Perbal B , Bergmann D G , Baluda M A
Published: January 2020

The avian myeloblastosis virus provirus inserted in a lambda bacteriophage, recombinant clone 11A1-1 (Souza et al., Proc. Natl. Acad. Sci. U.S.A. 77:3004-3008, 1980), was transfected into chicken embryo fibroblasts which had been preinfected with either Rous-associated virus type 61 or the transformation-defective avian sarcoma virus tdB77. Within 4 to 5 h after transfection, the cells were injected into 16-day-old chicken embryos or 1-day-old chicks. Acute myeloblastic leukemia developed after a long latent period. Filtered (0.22-micrometer pores) supernatant of transformed buffy-coat cell cultures from one leukemic chicken of the lambda 11A1-1 (tdB77) group rapidly transformed yolk sac cells in vitro. Results from an infectivity interference assay and analysis of proviral DNA fragments generated with restriction endonucleases were consistent with the presence in leukemic cells of defective avian myeloblastosis virus and tdB77 as the helper virus.
Normal translation of human adenovirus mRNA in cell-free lysates prepared from abortively as well as productively infected monkey cells
Authors: Quinlan M P , Klessig D F
Published: January 2020

A variety of mRNAs, including adenovirus-specified mRNAs isolated from infected human cells, were translated with similar efficiencies in S10 cell-free lysates prepared from productively and abortively infected monkey cells. These results may suggest that reduced synthesis of the late viral proteins in abortively infected monkey cells is not due to a defect in the protein-synthesizing apparatus of the cell.
Structure and biochemical functions of four simian virus 40 truncated large-T antigens
Authors: Chaudry F , Harvey R , Smith A E
Published: January 2020

The structure of four abnormal T antigens which are present in different simian virus 40 (SV40)-transformed mouse cell lines was studied by tryptic peptide mapping, partial proteolysis fingerprinting, immunoprecipitation with monoclonal antibodies, and in vitro translation. The results obtained allowed us to deduce that these proteins, which have apparent molecular weights of 15,000, 22,000, 33,000 and 45,000, are truncated forms of large-T antigen extending to different amounts into the amino acid sequences unique to large-T. The proteins are all phosphorylated, probably at a site between amino acids 106 and 123. The mRNAs coding for the proteins probably contain the normal large-T splice but are shorter than the normal transcripts of the SV40 early region. The truncated large-Ts were tested for the ability to bind to double-stranded DNA-cellulose. This showed that the 33,000- and 45,000-molecular-weight polypeptides contained sequences sufficient for binding under the conditions used, whereas the 15,000- and 22,000-molecular-weight forms did not. Together with published data, this allows the tentative mapping of a region of SV40 large-T between amino acids 109 and 272 that is necessary and may be sufficient for the binding to double-stranded DNA-cellulose in vitro. None of the truncated large-T species formed a stable complex with the host cell protein referred to as nonviral T-antigen or p53, suggesting that the carboxy-terminal sequences of large-T are necessary for complex formation.
Replication of mouse hepatitis virus: negative-stranded RNA and replicative form RNA are of genome length
Authors: Lai M M , Patton C D , Stohlman S A
Published: January 2020

There are seven virus-specific mRNA species in mouse hepatitis virus-infected cells (Lai et al., J. Virol. 39:823-834, 1981). In this study, we examined virus-specific negative-stranded RNA to determine whether there are corresponding multiple negative-stranded RNAs. Intracellular RNA from mouse hepatitis virus-infected cells was separated by agarose gel electrophoresis, transferred to nitrocellulose membranes, and hybridized to positive-stranded genomic 60S [32P]RNA. Only a single RNA species of genomic size was detected under these conditions. This RNA was negative stranded. No negative-stranded subgenomic RNA was detected. We also studied double-stranded replicative-form RNA in the infected cells. Only one replicative-form of genomic size was detected. When the double-stranded RNA isolated without RNase treatment was analyzed, again only one RNA species of genomic size was detectable. Furthermore, most of the virus-specific mRNAs could be released from this RNA species upon heating. These results suggest that all of the mouse hepatitis virus-specific RNAs are transcribed from a single species of negative-stranded RNA template of genomic size.
Effect of aphidicolin on avian sarcoma virus replication
Authors: Hsu T W , Taylor J M
Published: January 2020

We studied the effect of aphidicolin, an inhibitor of eucaryotic DNA polymerase alpha, on viral DNA replication and integration during the first 24 h after infection of quail embryo fibroblasts with avian sarcoma virus. In drug-treated cells, the synthesis of unintegrated linear viral DNA species was not impaired; however, the subsequent accumulation of circular viral DNA species and integrated proviral DNA was reversibly inhibited. After removal of the drug, circular viral DNA species were derived from preexisting linear viral DNA species, instead of being derived by de novo synthesis.
Homologous Terminal Sequences in the Double-Stranded RNA Genome Segments of Cytoplasmic Polyhedrosis Virus of the Silkworm
Bombyx mori
Authors: Kuchino Yoshiyuki , Nishimura Susumu , Smith Robert E. , Furuichi Yasuhiro
Published: January 2020

The 3′-terminal regions (20 to 32 residues) of the genome double-stranded RNA (dsRNA) segments of cytoplasmic polyhedrosis virus were sequenced. The dsRNAs, which were labeled at their 3′ termini by incubation with [5′-
32
P]pCp and T4 RNA ligase, were denatured and resolved into the plus and minus strands by agarose-urea gel electrophoresis. Ten single-stranded RNAs thus obtained from the five dsRNA segments IV, V, VIII, IX, and X were sequenced by postlabeling methods. Common 3′-terminal sequences, -GUUAGCC and -UUACU, were found in the plus and minus strands, respectively, of all five dsRNA segments. However, adjacent sequences diverged and were considerably variable. The homologous sequences found in the 3′ end may be important recognition signals for viral RNA polymerases and for assembly of the genome segments.
Newcastle disease virus stimulates the cellular accumulation of stress (heat shock) mRNAs and proteins
Authors: Collins P L , Hightower L E
Published: January 2020

A biological agent, Newcastle disease virus, stimulated the synthesis of stress proteins in cultured chicken embryo cells. Previously, only physical and chemical agents were known to induce these proteins. The levels of translatable stress mRNAs were elevated in cells infected with avirulent or virulent strains; however, stress protein synthesis was stimulated strongly only in cells infected by avirulent strains. As did several other paramyxoviruses, avirulent strains of Newcastle disease virus stimulated the synthesis of glucose-regulated proteins as well as stress proteins. Possible stimuli of the synthesis of these two sets of proteins in paramyxovirus-infected cells are considered.
Restriction enzyme analysis of partially transformation-defective mutants of acute leukemia virus MC29
Authors: Enrietto P J , Hayman M J
Published: January 2020

Restriction enzyme mapping and limited sequence analysis have been used to study the generation and genome structure of three partial-transformation mutants of avian acute leukemia virus MC29. The three mutants, td10A, td10C, and td10H, could be shown to have sustained overlapping deletions of 200, 400, and 600 base pairs, respectively, in their genomes. The precise location of the deletions was mapped within the v-myc gene of the mutants by limited sequence analysis of cloned MC29 DNA. The data obtained are discussed in terms of the effect of these deletions on the mechanism of transformation by MC29.
Avian sarcoma virus gag precursor polypeptide is not processed in mammalian cells
Authors: Vogt V M , Bruckenstein D A , Bell A P
Published: January 2020

We studied intracellular avian gag proteins (internal structural proteins of virions) in several mammalian cell lines transformed by Rous sarcoma virus. All lines examined contain gag antigens as determined by radioimmune assay. We used the techniques of protein blotting from polyacrylamide gels, which detects nanogram quantities of viral protein, to investigate the size of intracellular viral polypeptides. All of the lines that contained enough viral protein to be amenable to this type of analysis synthesized Pr76, the avian sarcoma virus gag precursor polypeptide, but failed to process it into mature virion proteins. In some cell lines, the recovery of Pr76 was greatly enhanced by the addition of a mixture of protease inhibitors, including the sulfhydryl-blocking reagent N-ethylmaleimide, to the lysis buffer. At least several of the mammalian cells also synthesized a viral polypeptide the size of Pr180, the precursor to reverse transcriptase. Since Rous sarcoma virus does not replicate or replicates extremely poorly in mammalian cells, the lack of processing suggests that cleavage and virion assembly are invariably associated.
Protease bypass of temperature-sensitive murine leukemia virus maturation mutants
Authors: Traktman P , Baltimore D
Published: January 2020

Cells infected with certain temperature-sensitive mutants of Moloney murine leukemia virus synthesize the virion precursor proteins but neither bud virions nor cleave precursor proteins to their mature form. Addition of proteases to cells infected with these mutants caused cleavage of the precursor proteins Pr65gag and Pr180gag-pol to their mature forms at the nonpermissive temperature. Concomitantly there was release from the cells of morphologically normal virions. The enzymatically inactive Pr180gag-pol was cleaved to active reverse transcriptase (p85), which was found in the released particles. External protease treatment apparently bypassed the lesion in these viral mutants, suggesting that their defect may involve a virus-specific protease.
mlt
Mutants of Polyoma Virus
Authors: Ding DaMing , Dilworth S. M. , Griffin Beverly E.
Published: January 2020

New
mlt
deletion mutants of polyoma virus were isolated, and their abilities to produce a lytic response in mouse cells or to transform rat cells were assessed. Their properties were analyzed in terms of the sequences deleted and their effects upon the structure and functions of the viral middle and large T-antigens.
Two Unique RNA Species of the Nodavirus Black Beetle Virus
Authors: Clewley Jonathan P. , Crump W. Andrew L. , Avery Roger J. , Moore Norman F.
Published: January 2020

Two-dimensional fingerprinting of RNase T
1
-derived oligonucleotides of the two individual RNA segments of the Nodavirus black beetle virus indicates that each RNA species possesses a distinct nucleotide sequence. Species 1 RNA has a genome complexity of approximately 3,000 nucleotides, and species 2 RNA is composed of approximately 1,500 nucleotides. Submolar amounts of oligonucleotides apparently derived from a third virus-specific RNA were also detected in black beetle virus RNA preparations.
Coronavirus proteins: biogenesis of avian infectious bronchitis virus virion proteins
Authors: Stern D F , Sefton B M
Published: January 2020

We examined the synthesis of viral structural proteins in cultured cells infected with the avian coronavirus infectious bronchitis virus. Tryptic peptide mapping was used to determine the structural relationships of the intracellular proteins to the virion polypeptides. Pulse-chase experiments were performed to identify precursors to the virus-specific proteins. We found that the nucleocapsid protein, P51, and the small viral membrane proteins GP31, GP28, and P23 do not undergo post-translational proteolytic processing. In contrast, GP90 and GP84, the two large virion membrane proteins, were found to be produced by cleavage of a single precursor, GP155. This demonstrated that at least one coronavirus mRNA specifies two virion proteins.
Association of viral particles and viral proteins with membranes in SA11-infected cells
Authors: Soler C , Musalem C , Loroño M , Espejo R T
Published: January 2020

Electron microscopy after negative staining of SA11-infected cell homogenates revealed that most of the viral particles are associated with membrane-like material. Many of the particles seemed to be fully enveloped in a membrane. This association could also be detected by the observed cosedimentation of viral proteins and cell membranes. Pulse-chase experiments showed that viral glycoproteins rapidly associate with membranes, whereas most of the structural proteins appearing in the soluble fraction immediately after the pulse were slowly chased into the membrane fraction. The membranes could be further fractionated into at least four fractions differing in density and containing a different distribution of viral proteins. Also, the distribution of label into each of these membrane fractions changed after long chase periods. The inhibition of glycosylation with tunicamycin yielded viral particles without an outer layer, but did not affect the described association with membranes. The possible relationship of this finding to the maturation of the virion is discussed.
Nucleotide sequence of the envelope gene of Friend murine leukemia virus
Authors: Koch W , Hunsmann G , Friedrich R
Published: January 2020

The envelope gene of the helper-independent, highly leukemogenic virus Friend murine leukemia virus was sequenced by using a molecular clone of a Friend murine leukemia provirus. The deduced amino acid sequences of the envelope proteins gp70 and p15env were homologous to the sequences of Moloney murine leukemia virus (86%) and Akv (76%). However, a stretch of about 40 amino acid residues near the middle of gp70 was dissimilar in Friend and Moloney murine leukemia viruses and Akv. In this type-specific region the gp70s of all three viruses contained more than 30% proline residues, giving this sequence a very rigid conformation. We suggest that this rigid and highly variable region of gp70 participates in infection by recognition of cell surface receptors and, in addition, might contribute to the different oncogenic spectra of murine leukemia viruses.
Integration of New Endogenous Mouse Mammary Tumor Virus Proviral DNA at Common Sites in the DNA of Mammary Tumors of C3Hf Mice and Hypomethylation of the Endogenous Mouse Mammary Tumor Virus Proviral DNA in C3Hf Mammary Tumors and Spleens
Authors: Etkind Polly R. , Sarkar Nurul H.
Published: January 2020

To understand the molecular mechanisms by which the endogenous murine mammary tumor virus (MuMTV) proviruses are expressed and produce late-occurring mammary tumors in C3Hf mice, we analyzed, by the use of restriction enzymes and the Southern transfer procedure, genomic DNA from normal organs of mammary tumor-bearing and tumor-free mice and from 12 late-occurring C3Hf mammary tumors. We found, by using the restriction enzymes
Eco
RI and
Hind
III, that in addition to the preexisting endogenous MuMTV proviruses, new MuMTV-specific proviral DNA was integrated into new sites in the host genome in all 12 of the tumors that we examined.
Pst
I digests of C3Hf tumor DNA revealed that the new proviral DNA found in C3Hf tumors was of endogenous origin. Moreover, the respective sizes of at least one of the new DNA fragments generated by
Eco
RI or
Hind
III digestion were the same in at least 50% of the C3Hf tumors analyzed, suggesting that the integration site of this new proviral DNA could be at the same location in the host genome of these tumors. Our results may imply that mammary tumorigenesis in C3Hf mice results from activation of cellular oncogenes by an MuMTV proviral DNA promoter. Specific hypomethylation of MuMTV proviral DNA was detected in the mammary tumors and spleens of C3Hf tumor-bearing mice. Our results indicated that most, if not all, of the hypomethylated MuMTV proviral DNA sequences were derived from the endogenous MuMTV provirus located at the MTV-1 locus, a locus responsible for the production of MuMTV antigens and increased incidence of mammary carcinoma in C3Hf mice. In spleens of non-tumor-bearing mice of ages 3, 6, 9, and 12 months, there was progressive hypomethylation of proviral DNA with increasing age, suggesting a possible correlation between demethylation of MuMTV proviral DNA in the spleens of C3Hf mice and the expression of endogenous MuMTV.
Physical Analysis of
Autographa californica
Nuclear Polyhedrosis Virus Transcripts for Polyhedrin and 10,000-Molecular-Weight Protein
Authors: Smith Gale E. , Vlak Just M. , Summers Max D.
Published: January 2020

In
Autographa californica
nuclear polyhedrosis virus-infected cells, polyhedrin, the major structural polypeptide of the viral occlusions, and a low-molecular-weight viral protein with a molecular weight of approximately 10,000 (10K) accumulated to high levels late in infection. Two polyadenylated RNAs 1,200 and 630 bases in size were the most abundant viral transcripts present in the cytoplasm at 48 h postinfection. Evidence is presented that these RNAs were the transcripts for polyhedrin and 10K proteins. The 630-base RNA and five other major polyadenylated RNAs present at 48 h postinfection mapped within or near
A. californica
nuclear polyhedrosis virus
Eco
RI-P. The DNA sequences that code for polyhedrin mRNA were examined by S1 nuclease analysis. The polyhedrin gene contained no detectable introns and mapped at 3,990 to 5,200 base pairs to the right of the origin of the circular
A. californica
nuclear polyhedrosis virus physical map. Heterogeneity at the 5′ end of polyhedrin mRNA was observed by using S1 nuclease analysis with 5′ end-labeled DNA probes.
Polypeptide structure and encoding location of the adenovirus serotype 2 late, nonstructural 33K protein
Authors: Oosterom-Dragon E A , Anderson C W
Published: January 2020

Radiochemical microsequence analysis of selected tryptic peptides of the adenovirus type 2 33K nonstructural protein has revealed the precise region of the genomic nucleotide sequence that encodes this protein. The initiation codon for the 33K protein lies 606 nucleotides to the right of the EcoRI restriction site at 70.7 map units and 281 nucleotides to the left of the postulated carboxyterminal codon of the adenovirus 100K protein. The coding regions for these two proteins thus overlap; however, the 33K protein is derived from the +1 frame with respect to the postulated 100K reading frame. Our results contradict an earlier published report suggesting that these two proteins share extensive amino acid sequence homology (N. Axelrod, Virology 87:366-383, 1978). The published nucleotide sequence of the Ad2 EcoRI-F fragment (70.7 to 75.9 map units) cannot accommodate in a single reading frame the peptide sequences of the 33K protein that we have determined. Sequence analysis of DNA fragments derived from virus has confirmed the published nucleotide sequence in all critical regions with respect to the coding region for the 33K protein. Consequently, our data are only consistent with the existence of an mRNA splice within the coding region for 33K. Consensus donor and acceptor splice sequences have been located that would predict the removal of 202 nucleotides from the transcripts for the 33K protein. Removal of these nucleotides would explain the structure of a peptide that cannot otherwise be directly encoded by the EcoRI-F fragment. Identification of the precise splice points by peptide sequencing has permitted a prediction of the complete amino acid sequence for the 33K protein.
Characterization of the 5' termini of purified nascent simian virus 40 late transcripts
Authors: Lycan D E , Danna K J
Published: January 2020

The primary transcripts of simian virus 40 are extensively processed in the nuclei of infected monkey cells before they are transported to the cytoplasm as mature mRNAs. To investigate the early steps in this process, in particular, to determine which events occur on nascent chains before the termination of transcription, we have developed a procedure for the purification of nascent viral transcripts. This technique involves the in vitro incorporation of mercurated residues into the growing 3' ends of pre-initiated nascent chains, allowing their specific purification by sulfhydrylcellulose affinity chromatography. We further selected viral specific transcripts by hybridization to simian virus 40 DNA-cellulose. We describe here our analysis of the 5' termini of purified nascent simian virus 40 transcripts. This analysis revealed various cap structures, providing direct evidence that primary viral transcripts are capped before chain completion. The various cap structures exhibited a full range of methylation states. Completely unmethylated GpppA cap cores were identified, as well as caps methylated at the penultimate position only. The presence of GpppAm and GpppmAm caps indicates that, in BSC-1 cells, the penultimate nucleotide can be methylated before 7-methyl-G formation. Furthermore, the proportions of the various intermediates suggest that, in contrast to the viral capping enzymes of vaccinia virus and reovirus, the cellular enzymes methylate in the following order: GpppA leads to GpppAm leads to GpppmAm leads to 7mGpppmAm. In addition to capped ends, we also detected some unprocessed pppA ends. To our knowledge, this is the first time uncapped termini have been identified on RNAs known to be polymerase II products.
Nucleotide sequences of feline sarcoma virus long terminal repeats and 5' leaders show extensive homology to those of other mammalian retroviruses
Authors: Hampe A , Gobet M , Even J , Sherr C J , Galibert F
Published: January 2020

The nucleotide sequences of the Gardner-Arnstein feline sarcoma virus (FeSV) long terminal repeat and the adjacent leader sequences 5' to the viral gag gene were determined. These were compared with homologous portions of Synder-Theilen FeSV and with previously published sequences for Moloney murine sarcoma virus and simian sarcoma virus proviral DNA. More than 75% of the residues in the FeSV R and U5 regions were homologous to sequences within the same regions of the other viral long terminal repeats. Unexpectedly, alignment of the FeSV sequences with those of the Moloney murine sarcoma and simian sarcoma viruses showed similar extents of homology within U3. The homologous U3 regions included the inverted repeats, a single set of putative enhancer sequences, corresponding to a "72-base-pair" repeat, and sequences, including the CAT and TATA boxes, characteristic of eucaryotic promotors. The 5' leader sequences of both FeSV strains included a binding site for prolyl tRNA and a putative splice donor sequence. In addition, the FeSV leader contained a long open reading frame which was adjacent to and in phase with the ATG codon at the 5' end of the FeSV gag gene. The open reading frame could code for a signal peptide of about 7.4 kilodaltons. Our results support the concept that the virogenic portions of both FeSV and simian sarcoma virus were ancestrally derived from viruses of rodent origin, with conservation of regulatory sequences as well as the viral structural genes.
Organization and stability of endogenous xenotropic murine leukemia virus proviral DNA in mouse genomes
Authors: Hoggan M D , Buckler C E , Sears J F , Rowe W P , Martin M A
Published: January 2020

In a series of blot hybridization experiments, using a xenotropic envelope probe and restriction enzymes known to cut xenotropic proviral DNA a single time (EcoRI) or not at all (HindIII), we have studied the organization and relationship of endogenous xenotropic env-related sequences in various mouse strains. Multiple copies (18 to 28) of xenotropic env-reactive fragments were found in all mouse DNAs after digestion with either HindIII or EcoRI, and the majority of fragments were of sizes compatible with their origin from full-length proviral DNA. Five HindIII and five EcoRI restriction fragments were common to all inbred mouse DNAs tested. In addition, each strain exhibited unique characteristic xenotropic env-reactive bands; these bands were remarkably stable during many years of inbreeding. The cleavage patterns characteristic of each strain were also useful for showing genealogical relatedness among the various inbred mice.
Simian virus 40 promoters direct expression of the tetracycline gene in plasmid pACYC184
Authors: Jenkins F J , Howett M K , Rapp F
Published: January 2020

Insertion of HindIII DNA fragments into the HindIII site of plasmid pACYC184 destroys the promoter of the plasmid tetracycline resistance gene and causes Escherichia coli cells harboring recombinant plasmids to be tetracycline sensitive and chloramphenicol resistant. The HindIII-C DNA fragment of simian virus 40 contains the two virus promoters and the virus origin of replication. We report the isolation of recombinant plasmids that contained the simian virus 40 HindIII-C DNA fragment at the HindIII site but were capable of conferring tetracycline resistance to E. coli cells. The viral promoter sequences contained in the HindIII-C fragment presumably replaced the inactivated tetracycline resistance gene promoter sequences and enabled transcription of the tetracycline resistance gene.
Analysis of recombinant DNA clones of the endogenous BALB/c murine leukemia virus WN1802N: variation in long terminal repeat length
Authors: Boone L R , Myer F E , Yang D M , Kiggans J O , Koh C , Tennant R W , Yang W K
Published: January 2020

We analyzed 15 recombinant DNA clones of the unintegrated closed circular DNA intermediate of the BALB/c endogenous ecotropic murine leukemia virus WN1802N. Thirteen of these clones had an insert which corresponded to the complete murine leukemia virus genome. Of these, six contained a single long terminal repeat (LTR) and seven contained two LTRs. The viral genomes in nine clones had an LTR of 520 base pairs (bp), one had an LTR of 570 bp, three had an LTR of 600 bp, and one had an LTR of 670 bp. Restriction endonuclease analysis demonstrated that the size variability resides in the U3 region. Seven of eight clones which yielded infectious virus by DNA transfection had the 520-bp LTR, and the other had a 600-bp LTR. More detailed examination of plasmid subclones of three isolates with different-sized LTRs revealed that the approximate position which varies in the U3 region corresponds to the 72-bp repeat region of Moloney sarcoma virus. Possible consequences of these variations are discussed.
Vaccinia virus thymidine kinase and neighboring genes: mRNAs and polypeptides of wild-type virus and putative nonsense mutants
Authors: Bajszár G , Wittek R , Weir J P , Moss B
Published: January 2020

Enzymatically active thymidine kinase (TK) was made in reticulocyte lysates programmed with early vaccinia mRNA that hybridized to plasmid recombinants containing either of two adjacent small DNA subsegments of the viral HindIII-J fragment. The map position of an early polypeptide, with a molecular weight of 19,000 (19K), coincided precisely with that of the TK. The absence of the 19K polypeptide in cell-free translation products of hybridization-selected mRNAs from several TK-negative mutants provided an independent identification of the TK polypeptide. The small size of the TK polypeptide of vaccinia virus distinguishes it from that of procaryotes, eucaryotes, and herpesvirus. Five early mRNAs of 3,840, 2,390, 1,790, 1,070, and 590 nucleotides were mapped within the HindIII-J fragment by RNA blotting and nuclease S1 digestion of RNA-DNA hybrids. The RNAs of 590 and 2,380 nucleotides were found to have 5' coterminal ends and represent major and minor forms, respectively, of the TK message. The 3' end of the minor TK mRNA appeared to be coterminal with the 3' end of the 1,790-nucleotide transcript which encodes a 41K polypeptide. The 1,070-nucleotide RNA was identified as the message for a 21K polypeptide. All of these RNAs, including the two forms of the TK message, were made by the putative TK-negative nonsense mutants.
Mapping of a 14,000-dalton antigen to early region 4 of the human adenovirus 5 genome
Authors: Downey J F , Rowe D T , Bacchetti S , Graham F L , Bayley S T
Published: January 2020

An early 14,000-dalton (14K) antigen of adenovirus 5, hitherto designated 10.5K and thought to be from early region 1 (E1), has been shown to be a product of region E4 on the following evidence. In KB cells infected with the adenovirus 5 mutants dl312 and dl313, containing large deletions in region E1, this antigen was produced in a form having the same mobility as that in wild-type infections. In a range of rodent cells transformed by adenovirus 5 DNA, the synthesis of 14K antigen and the ability of these cells to elicit an immune response to this protein both correlated with the presence of sequences from region E4 of the viral genome. A 14K polypeptide was synthesized in a cell-free system programmed with infected-cell mRNA and was found to be identical to the in vivo antigen in antigenicity, in electrophoretic mobility, and in [35S]methionine-containing tryptic peptides. After labeling with [35S]methionine and several 3H-amino acids, this in vitro product gave an N-terminal sequence identical to that expected from one of the open reading frames (reading region 3) in the DNA sequence for region E4 of Hérissé et al. (Nucleic Acids Res. 9:4023-4042, 1981). It is likely that this antigen is the same as the nucleus-associated 11K polypeptide from E4 described by other authors.
Restriction endonuclease map of corynebacteriophage omega ctox+ isolated from the Park-Williams no. 8 strain of Corynebacterium diphtheriae
Authors: Rappuoli R , Michel J L , Murphy J R
Published: January 2020

The toxigenic corynebacteriophage omega tox+ was isolated from the hypertoxigenic Park-Williams no. 8 (PW8) strain of Corynebacterium diphtheriae and compared with the toxigenic corynebacteriophage beta tox+. The physical size and host range of both phages were found to be identical. An endonuclease restriction map of omega tox+ was constructed, and the locations of the cohesive ends (cos), phage attachment site (attP), and the diphtheria tox operon were identified. The genome of omega tox+ was found to differ from that of beta tox+ in three regions. In addition, omega tox+ was shown to be integrated into two nontandem corynebacterial phage attachment sites (attB1, attB2) in the PW8 chromosome. The differences in the restriction endonuclease digestion maps of omega tox+ and beta tox+ and the contribution of double lysogeny are discussed in relation to the hypertoxigenicity of the PW8 strain.
Isolation and characterization of a membrane-bound population of group B coxsackieviruses
Authors: Chatterjee N K , Samsonoff W A , Tuchowski C
Published: January 2020

HeLa cells infected with several group B coxsackieviruses contain a previously undetected, virus-specific ribonucleoprotein particle which we designated membrane-bound virion (MBV). MBVs of B5 virus have a pronounced polygonal appearance and are slightly smaller than virions. The particles sediment more slowly (at about 107S) and have a lower buoyant density (about 1.30). They contain 35S virion RNA; only three, and not four, capsid proteins; and at least seven additional proteins with apparent molecular weights of 21,000 to 92,000. Three of the latter proteins appear to be of host origin; the rest may be precursors of virion capsid proteins. The RNA is resistant to digestion by RNase, and EDTA treatment disrupts the particle. MBVs are infectious, although significantly less so than virions. Cells infected with MBVs produce both types of progeny, virions and MBVs. In coinfected cultures, the yield of progeny is lower than in cells infected with virions alone, suggesting interference by MBVs. Synthesis of both types can be detected within 3.5 h after infection, and synthesis continues for 24 h.
Additional Restriction Endonuclease Cleavage Sites on the Bacteriophage P22 Genome
Authors: Casjens Sherwood , Hayden Melody , Jackson Ethel , Deans Robert
Published: January 2020

We present complete restriction endonuclease cleavage site maps of the bacteriophage P22 chromosome for 16 enzymes with six base recognition sequences, thereby positioning 116 new sites on the chromosome. Twenty-four such restriction maps for P22 DNA, containing 162 sites, have now been completed, and three enzymes were found that did not cut P22 DNA. Our results are consistent with the ideas that
Cla
I does not cleave the methylated recognition sequence ATCGA
me
T or A
me
TCGAT and
Stu
I does not cleave the methylated recognition sequence AGGCC
me
T.
Nucleotide Sequence of the Long Terminal Repeat and Flanking Cellular Sequences of Avian Endogenous Retrovirus
ev
-2: Variation in Rous-Associated Virus-0 Expression Cannot Be Explained by Differences in Primary Sequence
Authors: Scholl D. R. , Kahn S. , Malavarca R. , Astrin S. , Skalka A. M.
Published: January 2020

A fragment of chicken DNA containing the left long terminal repeat of endogenous retrovirus
ev
-2 and flanking cellular sequences has been molecularly cloned and analyzed. Comparison with sequence data from the analogous regions of
ev
-1 and Rous-associated virus-0 viral DNA reveals similarities among flanking regions of the integrated proviruses and among all three long terminal repeats. From the latter finding, we conclude that the difference in level of expression of
ev
-2 and its progeny Rous-associated virus-0 provirus cannot be due to sequence differences in their upstream long terminal repeats.
Cyanogen bromide digestion of the avian myeloblastosis virus pp19 protein: isolation of an amino-terminal peptide that binds to viral RNA
Authors: Johnson S P , Veigl M , Vanaman T , Leis J
Published: January 2020

The avian myeloblastosis virus pp19 protein was separated from the other virus proteins by a rapid and simple purification procedure which yields milligram amounts of homogeneous protein. This protein was then fragmented by digestion with cyanogen bromide. When the mixture of the cyanogen bromide peptides was passed through a 60S avian myeloblastosis virus RNA-cellulose column, only one peptide bound with high affinity to the resin. The peptide migrated on a sodium dodecyl sulfate-polyacrylamide gel with an approximate molecular weight of 2,900 and will be referred to as the p3B peptide. This peptide was also isolated directly by chromatography of the cyanogen bromide-digested pp19 protein on a reverse-phase high-pressure liquid chromatography column. It was again the only cyanogen bromide peptide of the pp19 protein that bound to the RNA affinity resin. The p3B peptide is a basic peptide, as was seen by its rapid migration on acid-urea-polyacrylamide gels and its amino acid composition. A partial amino acid sequence analysis of the p3B peptide indicated that it was derived from the amino terminus of the intact protein. Although the p3B peptide bound to 60S RNA, it did not demonstrate the selective binding of native pp19 to regions of the RNA containing secondary structure.
Derivation of Neutralizing Monoclonal Antibodies Against Rotavirus
Authors: Sonza Secondo , Breschkin Alan M. , Holmes Ian H.
Published: January 2020

Monoclonal antibodies were derived against the SA11 simian, NIC bovine, and Wa human rotavirus strains and characterized by enzyme-linked immunosorbent assay, plaque neutralization, and hemagglutination inhibition. Several strain SA11-specific antibodies were found to have neutralizing and hemagglutination-inhibiting capacity.
Comparison of the sequences and coding of La Crosse and snowshoe hare bunyavirus S RNA species
Authors: Akashi H , Bishop D H
Published: January 2020

The sequence of the S RNA of La Crosse bunyavirus was deduced from analyses of DNA copies cloned in the Escherichia coli plasmid pBR322. The S RNA is 984 nucleotides in length, has a base ratio of 31.8% U, 27.0% A, 23.2% C, and 18.0% G, and codes for two distinct gene products that are read from overlapping reading frames in the viral complementary strand. The larger gene product (N, 26.5 x 10(3) daltons) contains 235 amino acids, and the smaller gene product (NSS, 10.4 x 10(3) daltons) has 92 amino acids. Comparisons with the published sequences of the related snowshoe hare bunyavirus S RNA and its gene products (Bishop et al., Nucleic Acids Res. 10:3703-3713, 1982) indicate that there are a total of 114 nucleotide differences (6 additions or deletions and 108 substitutions). Also, there are 22 amino acid differences between the N proteins and 12 amino acid differences between the NSS proteins of the two S RNAs.
P85: a
gag-mos
Polyprotein Encoded by
ts
110 Moloney Murine Sarcoma Virus
Authors: Stanker L. H. , Gallick G. E. , Kloetzer W. S. , Murphy E. C. , Arlinghaus R. B.
Published: January 2020

Antibody to a synthetic peptide (anti-C3 serum) with the predicted sequence of the C terminus of the Moloney murine sarcoma virus (strain 124) v-
mos
gene was used in immunoprecipitation experiments with cytoplasmic extracts of a clone of NRK cells infected with
ts
110 Moloney murine sarcoma virus, termed 6m2 cells.
ts
110 Moloney murine sarcoma virus codes for two viral proteins of 85,000 and 58,000
M
r
, termed P85 and P58, respectively, in nonproducer 6m2 cells maintained at 33°C. Anti-C3 serum specifically recognized [
3
H]leucine-labeled P85, but not P58, from infected cells maintained at 33°C, whereas antiserum prepared against murine leukemia virus p12 recognized both proteins. Normal serum and anti-C3 serum pretreated with excess C3 peptide did not precipitate P85. Immunoprecipitation experiments after metabolic labeling of 6m2 cells with
32
P
i
showed that P85 is phosphorylated. Both anti-C3 and anti-p12 sera specifically detected
32
P-labeled P85. Cell-free translation of
ts
110 murine sarcoma virus/murine lukemia virus RNA produces P85, P58, and helper virus protein Pr63

gag

. Anti-C3 serum specifically precipitated P85 but neither P58 nor Pr63

gag

. We conclude from these studies that P85 is a product of both the
gag
and
mos
genes of
ts
110 murine sarcoma virus, and, therefore, it is referred to as P85

gag-mos

. We have not detected any other v-
mos
gene product in
ts
110-infected cells.
Cloning and analysis of reverse transcript P160 genomes of Abelson murine leukemia virus
Authors: Latt S A , Goff S P , Tabin C J , Paskind M , Wang J Y , Baltimore D
Published: January 2020

Circular duplex reverse transcripts of the genome of a strain of Abelson murine leukemia virus that encodes a 160,000-molecular-weight protein were isolated, cleaved with HindIII restriction endonuclease, and cloned into the unique HindIII site of lambda phage Charon 21A. Recombinant phage clones, some of which were infectious in transfection assays, were found to contain a 789-base-pair region specific for Abelson murine leukemia virus; this region is not found in other strains of this virus. The extra sequence was localized by restriction endonuclease and electron microscopic heteroduplex analysis. Sequence analysis showed no homology at the ends of the extra sequence, implying that it was deleted by an event that did not utilize sequence homology. The sequence of this unique region has an open reading frame through its entirety.
Study of the Functional Activities Concomitantly Retained by the 115,000
M
r
Super T Antigen, an Evolutionary Variant of Simian Virus 40 Large T Antigen Expressed in Transformed Rat Cells
Authors: May Evelyne , Lasne Claude , Prives Carol , Borde Janine , May Pierre
Published: January 2020

Simian virus 40 (SV40) transformed V 11 F 1 clone 1 subclone 7 rat cells (subclone 7) do not synthesize normal-size large T antigen (

M
r

, 90,000); instead, they produce a 115,000

M
r

super T antigen (115K super T antigen). This super T antigen is SV40 virus coded, and its synthesis results from rearrangement and amplification of integrated viral DNA sequences in subclone 7 (May et al., Nucleic Acids Res.
9
:4111-4128, 1981). In this study the functional activities of 115K super T antigen were compared with the functional activities of SV40 large T antigen. Transfection experiments were performed with (i) cosmid SVE 5 Kb and plasmid pSVsT, both containing the super T antigen gene and (ii) plasmids pSV1 and pSV40, both containing the large T antigen gene. Transfection of pSVsT DNA or SVE 5 Kb DNA into secondary cultures of rat kidney cells induced the formation of transformed cell foci with an efficiency that was about 50% of the efficiency of pSV1 DNA or pSV40 DNA. Concomitant with the transforming activity, two other activities were also retained by super T antigen, namely, the ability to enhance the level of host cellular protein p53 and the capacity to bind to p53. In contrast, pSVsT and SVE 5 Kb DNAs were markedly deficient in the capacity to support
tsA
58 DNA replication in CV1-P cells at a nonpermissive temperature (41°C), as shown by cotransfection experiments. The yield of virus produced in these experiments was 400-fold less than the yield obtained in parallel experiments with pSV40 or pSV1. However, SVE 5 Kb and pSVsT have a functional SV40 replication origin, as shown by their efficient replication in COS 1 cells which provided functional large T antigen. Super T antigen also possesses a specific affinity for sequences of SV40 viral origin. Our results suggest that under certain conditions, evolutionary changes in T antigen take place and that these changes could be restricted to the phenotypic requirement of maintaining a structure that is able to induce cell transformation, to form a complex with p53, and to enhance the cellular level of p53. Therefore, there appears to be a close relationship among the activities of T antigen involved in transforming cells, in binding to p53, and in enhancing the p53 cellular level. Moreover, this set of activities appears to be separable from the replicative ability of T antigen, based on the observation that 115K super T antigen is markedly defective for initiating viral DNA synthesis.
Mouse immunoglobulin antibodies require complement for neutralization of mouse retroviruses
Authors: Sarma P S , Rowe W P
Published: January 2020

The addition of guinea pig complement was found to enhance the neutralizing capacity of mouse antibodies directed against the endogenous ecotropic murine leukemia viruses. The same immune sera, when tested without complement, had low or negligible neutralizing capacities, regardless of whether freshly harvested, unfrozen virus was used to preserve virus infectivity. Antibodies in high titers were found in sera from NFS congenic mice carrying the mouse leukemia virus inducing locus Akv-2. These mouse antibodies were type specific and failed to neutralize either Friend or Moloney leukemia virus. The mouse serum immunoglobulin fraction containing the complement-dependent antibodies was tentatively identified as immunoglobulin M.
Location of Homologous DNA Sequences Interspersed at Five Regions in the Baculovirus AcMNPV Genome
Authors: Cochran Mark A. , Faulkner Peter
Published: January 2020

An examination of
Autographa californica
nuclear polyhedrosis virus DNA revealed the presence of five interspersed regions, rich in
Eco
RI restriction sites, which shared homologous sequences. These homologous regions (hr), designated hr
1
to hr
5
, occur at or near the following
Eco
RI fragment junctions: hr
1
Eco
RI-B—
Eco
RI-I (0.0 map units); hr
2
,
Eco
RI-A—
Eco
RI-J (19.8 map units); hr
3
,
Eco
RI-C—
Eco
RI-G (52.9 map units); hr
4
,
Eco
RI-Q—
Eco
RI-L (69.8 map units); and hr
5
,
Eco
RI-S—
Eco
RI-X (88.0 map units). Four of these regions were identified, by cross-blot hybridization of
Hin
dIII-restricted
A. californica
nuclear polyhedrosis virus DNA, to be within the
Hin
dIII-A/B, -F, -L, and -Q fragments. The location of these regions and the identification of a fifth homologous region were confirmed, and their characterization was facilitated, by using two plasmids with
Hin
dIII-L or -Q fragment insertions, which contained the homologous regions hr
2
and hr
5
, respectively. The sizes of the homologous regions were about 800 base pairs for hr
2
, 500 base pairs for hr
5
, and less than 500 base pairs for hr
1
, hr
3
, and hr
4
. A set of small
Eco
RI fragments (
Eco
RI minifragments) which ranged in size from 225 to 73 base pairs were detected in
A. californica
nuclear polyhedrosis virus DNA and
Hin
dIII-L and -Q fragments by polyacrylamide gel analysis. Some of the minifragments in viral DNA were present in extramolar amounts and corresponded in size to some of the minifragments present in
Hin
dIII-L and -Q. Clones of some of the
Eco
RI minifragments were used as probes in hybridizations to digests of viral DNA and of
Hin
dIII-L and -Q. The hybridization data, obtained under various levels of stringency, suggested that there was a degree of mismatching between the sequences which were responsible for the homology.
Role of R loops in recA-independent homologous recombination of bacteriophage lambda
Authors: Matsumoto T , Ikeda H
Published: January 2020

We have previously shown that the DNA-dependent RNA polymerase of Escherichia coli can promote homologous recombination of bacteriophage lambda independently of the recA function. To detect this recombination, we jointly infected the cells with a pair of lambda phages in the presence of chloramphenicol, extracted intracellular lambda DNA molecules, packaged them in vitro, and measured the number of resulting recombinant phage particles. We showed that the recombination of DNA molecules takes place in vitro after extraction of DNA from the cells. We fractionated recombinogenic forms of intracellular lambda DNA and showed that they carry RNA. These and other results suggest that the R (RNA) loop structures, generated by transcription within the cells, promote homologous recombination in vitro. We discuss possible mechanisms of this recombination and compare those with the other forms of general recombination.
Structure and transcription of the cellular homolog (c-myb) of the avian myeloblastosis virus transforming gene (v-myb)
Authors: Gonda T J , Bishop J M
Published: January 2020

We isolated and characterized molecular clones containing the chicken cellular homolog (c-myb) of the avian myeloblastosis virus oncogene (v-myb). Mapping of the c-myb clones using restriction endonucleases and hybridization to radiolabeled v-myb probes revealed that the sequences homologous to v-myb are contained within four separate regions, which have since been shown by nucleotide sequencing (Klempnauer et al., Cell 31:453-463, 1982) to carry seven exons. Analysis of c-myb transcripts showed the presence of several large precursor RNAs in addition to the 4.0-kilobase cytoplasmic mRNA. We also determined the approximate positions of the c-myb sequences that are present in the 4.0-kilobase c-myb mRNA but not present in v-myb. Some of these sequences are found in a separate region 5' to the v-myb-related sequences, whereas the remainder appear to be located immediately 3' to the v-myb-related sequences. The data presented here, in conjunction with nucleotide sequence analysis (Klempnauer et al., Cell 31:453-463, 1982), indicate that the c-myb gene contains at least eight exons which span a total of about 16 kilobase pairs. The presence of exon sequences in c-myb outside the regions of homology with v-myb raises the possibility that the v-myb and c-myb gene products may differ significantly.