Detection of Avian Leukosis Viruses in Chickens by an in Vivo Rous Sarcoma Virus Genome Rescue Test

1971 ◽  
Vol 15 (2) ◽  
pp. 212 ◽  
Author(s):  
D. Katz ◽  
A. Kohn
Keyword(s):  
Rous Sarcoma Virus ◽  
Virus Genome ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Avian Leukosis Viruses

Transcription of the Rous Sarcoma Virus Genome In Vitro and In Vivo

2015 ◽  
pp. 517-523
Author(s):  
J. M. Bishop ◽  
C. T. Deng ◽  
A. J. Faras ◽  
H. M. Goodman ◽  
R. R. Guntaka ◽  
...  
Keyword(s):  
Rous Sarcoma Virus ◽  
Virus Genome ◽  
Rous Sarcoma ◽  
Sarcoma Virus

Precise localization of m6A in Rous sarcoma virus RNA reveals clustering of methylation sites: implications for RNA processing

1985 ◽  
Vol 5 (9) ◽  
pp. 2298-2306
Author(s):  
S E Kane ◽  
K Beemon
Keyword(s):  
Rous Sarcoma Virus ◽  
Rna Splicing ◽  
Paper Electrophoresis ◽  
Virus Genome ◽  
Rous Sarcoma ◽  
Virus Rna ◽  
Dna Restriction Fragments ◽  
Sarcoma Virus ◽  
Dna Restriction ◽  
Layer Chromatography

N6-methyladenosine (m6A) residues are present as internal base modifications in most higher eucaryotic mRNAs; however, the biological function of this modification is not known. We describe a method for localizing and quantitating m6A within a large RNA molecule, the genomic RNA of Rous sarcoma virus. Specific fragments of 32P-labeled Rous sarcoma virus RNA were isolated by hybridization with complementary DNA restriction fragments spanning nucleotides 6185 to 8050. RNA was digested with RNase and finger-printed, and individual oligonucleotides were analyzed for the presence of m6A by paper electrophoresis and thin-layer chromatography. With this technique, seven sites of methylation in this region of the Rous sarcoma virus genome were localized at nucleotides 6394, 6447, 6507, 6718, 7414, 7424, and 8014. Further, m6A was observed at two additional sites whose nucleotide assignments remain ambiguous. A clustering of two or more m6A residues was seen at three positions within the RNA analyzed. Modification at certain sites was found to be heterogeneous, in that different molecules of RNA appeared to be methylated differently. Previous studies have determined that methylation occurs only in the sequences Gm6AC and Am6AC. We observed a high frequency of methylation at PuGm6ACU sequences. The possible involvement of m6A in RNA splicing events is discussed.

Functional domains of the pp60v-src protein as revealed by analysis of temperature-sensitive Rous sarcoma virus mutants

1984 ◽  
Vol 4 (8) ◽  
pp. 1508-1514
Author(s):  
A W Stoker ◽  
P J Enrietto ◽  
J A Wyke
Keyword(s):  
Rous Sarcoma Virus ◽  
Kinase Activity ◽  
Temperature Sensitive ◽  
Tyrosine Kinase Activity ◽  
Rous Sarcoma ◽  
Heat Labile ◽  
Src Gene ◽  
Sarcoma Virus

Four temperature-sensitive (ts) Rous sarcoma virus src gene mutants with lesions in different parts of the gene represent three classes of alteration in pp60src. These classes are composed of mutants with (i) heat-labile protein kinase activities both in vitro and in vivo (tsLA27 and tsLA29), (ii) heat-labile kinases in vivo but not in vitro (tsLA33), and (iii) neither in vivo nor in vitro heat-labile kinases (tsLA32). The latter class indicates the existence of structural or functional pp60src domains that are required for transformation but do not grossly affect tyrosine kinase activity.

Regulation of Rous sarcoma virus RNA splicing and stability

1988 ◽  
Vol 8 (11) ◽  
pp. 4858-4867 ◽  
Author(s):  
S Arrigo ◽  
K Beemon
Keyword(s):  
Rous Sarcoma Virus ◽  
Rna Splicing ◽  
Negative Regulator ◽  
Splice Donor ◽  
Acceptor Pair ◽  
Cis Acting ◽  
Rous Sarcoma ◽  
Donor And Acceptor ◽  
Sarcoma Virus

Only a fraction of retroviral primary transcripts are spliced to subgenomic mRNAs; the unspliced transcripts are transported to the cytoplasm for packaging into virions and for translation of the gag and pol genes. We identified cis-acting sequences within the gag gene of Rous sarcoma virus (RSV) which negatively regulate splicing in vivo. Mutations were generated downstream of the splice donor (base 397) in the intron of a proviral clone of RSV. Deletion of bases 708 to 800 or 874 to 987 resulted in a large increase in the level of spliced RSV RNA relative to unspliced RSV RNA. This negative regulator of splicing (nrs) also inhibited splicing of a heterologous splice donor and acceptor pair when inserted into the intron. The nrs element did not affect the level of spliced RNA by increasing the rate of transport of the unspliced RNA to the cytoplasm but interfered more directly with splicing. To investigate the possible role of gag proteins in splicing, we studied constructs carrying frameshift mutations in the gag gene. While these mutations, which caused premature termination of gag translation, did not affect the level of spliced RSV RNA, they resulted in a large decrease in the accumulation of unspliced RNA in the cytoplasm.

scholarly journals COMPARATIVE STUDIES IN ROUS SARCOMA WITH VIRUS, TUMOR CELLS, AND CHICK EMBRYO CELLS TRANSFORMED IN VITRO BY VIRUS

1962 ◽  
Vol 115 (1) ◽  
pp. 245-251 ◽  
Author(s):  
Robert M. Dougherty ◽  
Herbert R. Morgan
Keyword(s):  
Chick Embryo ◽  
Tumor Cells ◽  
Comparative Studies ◽  
Rous Sarcoma Virus ◽  
Rous Sarcoma ◽  
Transformed Fibroblasts ◽  
Embryo Cells ◽  
Sarcoma Virus

Chick embryo fibroblasts infected in vitro with Rous sarcoma virus have properties similar to tumor cells when injected into virus-immune chickens. When such virus-transformed fibroblasts are injected into normal chickens, they apparently participate in the production of tumors independent of their release of virus and are thus apparently malignant in vivo.

scholarly journals Regulation of Rous sarcoma virus RNA splicing and stability.

1988 ◽  
Vol 8 (11) ◽  
pp. 4858-4867 ◽  
Author(s):  
S Arrigo ◽  
K Beemon
Keyword(s):  
Rous Sarcoma Virus ◽  
Rna Splicing ◽  
Negative Regulator ◽  
Splice Donor ◽  
Acceptor Pair ◽  
Cis Acting ◽  
Rous Sarcoma ◽  
Donor And Acceptor ◽  
Sarcoma Virus

Only a fraction of retroviral primary transcripts are spliced to subgenomic mRNAs; the unspliced transcripts are transported to the cytoplasm for packaging into virions and for translation of the gag and pol genes. We identified cis-acting sequences within the gag gene of Rous sarcoma virus (RSV) which negatively regulate splicing in vivo. Mutations were generated downstream of the splice donor (base 397) in the intron of a proviral clone of RSV. Deletion of bases 708 to 800 or 874 to 987 resulted in a large increase in the level of spliced RSV RNA relative to unspliced RSV RNA. This negative regulator of splicing (nrs) also inhibited splicing of a heterologous splice donor and acceptor pair when inserted into the intron. The nrs element did not affect the level of spliced RNA by increasing the rate of transport of the unspliced RNA to the cytoplasm but interfered more directly with splicing. To investigate the possible role of gag proteins in splicing, we studied constructs carrying frameshift mutations in the gag gene. While these mutations, which caused premature termination of gag translation, did not affect the level of spliced RSV RNA, they resulted in a large decrease in the accumulation of unspliced RNA in the cytoplasm.

A physical map of the Rous sarcoma virus genome

1976 ◽  
Vol 100 (3) ◽  
pp. 293-318 ◽  
Author(s):  
J.M. Coffin ◽  
M.A. Billeter
Keyword(s):  
Rous Sarcoma Virus ◽  
Physical Map ◽  
Virus Genome ◽  
Rous Sarcoma ◽  
Sarcoma Virus

scholarly journals Localization of 9E3/CEF-4 in avian tissues: expression is absent in Rous sarcoma virus-induced tumors but is stimulated by injury.

1990 ◽  
Vol 110 (3) ◽  
pp. 581-595 ◽  
Author(s):  
M Martins-Green ◽  
M J Bissell
Keyword(s):  
Rous Sarcoma Virus ◽  
Physiological Role ◽  
Wound Response ◽  
Angiogenic Factor ◽  
Target Cells ◽  
Embryonic Cells ◽  
Normal Tissues ◽  
Rous Sarcoma ◽  
Sarcoma Virus

The avian gene 9E3/CEF-4, a member of the superfamily of genes that includes KC and gro, is expressed abundantly in exponentially growing cultures of chick embryo fibroblasts (CEFs) and at high levels in CEFs transformed with Rous sarcoma virus (RSV). The product of this gene is a secreted protein that has homologies and structural similarities to inflammatory mediators. The function of 9E3 is obscure and its expression in vivo has not yet been investigated. We studied by in situ hybridization and RNA blots the pattern of 9E3 mRNA distribution in the wings of normal, wounded, and RSV-infected newly hatched chicks. We found that the message for 9E3 is high in specific tissues in normal wings; whereas connective tissue, tendon, and bone express the gene, muscle fibers, endothelium, epidermis, and bone marrow do not. The distribution coincides with that of interstitial collagen. Wounding results in marked elevation of the mRNA within the granulation tissue formed during healing and in adjacent tissues, especially those showing neovascularization. Similar elevation of mRNA occurs immediately adjacent to RSV tumors but, surprisingly, the tumor tissue itself shows no detectable levels of this message. Cells explanted from the tumors and grown in culture also show no expression of 9E3, in marked contrast to the very high level found in similarly cultured RSV-transformed CEFs. These results show that there are intrinsic differences between transformed embryonic cells in tissue culture and RSV target cells in the hatched chick. However, the expression of the gene in the periphery of tumors leaves open the possibility that 9E3 may still be involved in RSV carcinogenesis. The abundant expression of 9E3 in normal tissues indicates that the product of this gene plays a normal physiological role in tissues growing by cell division, perhaps as a growth regulator. The elevated expression of 9E3 in areas of neovascularization, makes it possible that the product of this gene could act as an angiogenic factor. Finally, expression in conjunction with high collagen levels and in wounded tissues may point to a role in wound response and/or repair, possibly via alteration of extracellular matrix.

scholarly journals Molecular and Genetic Determinants of Rous Sarcoma Virus Integrase for Concerted DNA Integration

2003 ◽  
Vol 77 (11) ◽  
pp. 6482-6492 ◽  
Author(s):  
Roger Chiu ◽  
Duane P. Grandgenett
Keyword(s):  
Rous Sarcoma Virus ◽  
Dnase I ◽  
Dna Interactions ◽  
Viral Dna ◽  
Catalytic Core ◽  
Dna Integration ◽  
Rous Sarcoma ◽  
Protein Dna Interactions ◽  
Sarcoma Virus

ABSTRACT Site-directed mutagenesis of recombinant Rous sarcoma virus (RSV) integrase (IN) allowed us to gain insights into the protein-protein and protein-DNA interactions involved in reconstituted IN-viral DNA complexes capable of efficient concerted DNA integration (termed full-site). At 4 nM IN, wild-type (wt) RSV IN incorporates ∼30% of the input donor into full-site integration products after 10 min of incubation at 37°C, which is equivalent to isolated retrovirus preintegration complexes for full-site integration activity. DNase I protection analysis demonstrated that wt IN was able to protect the viral DNA ends, mapping ∼20 bp from the end. We had previously mapped the replication capabilities of several RSV IN mutants (A48P and P115S) which appeared to affect viral DNA integration in vivo. Surprisingly, recombinant RSV A48P IN retained wt IN properties even though the virus carrying this mutation had significantly reduced integrated viral DNA in comparison to wt viral DNA in virus-infected cells. Recombinant RSV P115S IN also displayed all of the properties of wt RSV IN. Upon heating of dimeric P115S IN in solution at 57°C, it became apparent that the mutation in the catalytic core of RSV IN exhibited the same thermolabile properties for 3′ OH processing and strand transfer (half-site and full-site integration) activities consistent with the observed temperature-sensitive defect for integration in vivo. The average half-life for inactivation of the three activities were similar, ranging from 1.6 to 1.9 min independent of the IN concentrations in the assay mixtures. Wt IN was stable under the same heat treatment. DNase I protection analysis of several conservative and nonconservative substitutions at W233 (a highly conserved residue of the retrovirus C-terminal domain) suggests that this region is involved in protein-DNA interactions at the viral DNA attachment site. Our data suggest that the use of recombinant RSV IN to investigate efficient full-site integration in vitro with reference to integration in vivo is promising.

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