The 30S Moloney sarcoma virus RNA contains leukemia virus nucleotide sequences (original) (raw)
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Sequence homology between Moloney murine sarcoma virus and Moloney leukemia virus RNA
Journal of Virology, 1976
The Moloney murine sarcoma-leukemia virus [M-MSV (MuLV)], propagated at high multiplicity of infection (MOI), was demonstrated previously to contain a native genome mass of 4 X 10(6) daltons as contrasted to a mass of 7 X 10(6) daltons for Moloney murine leukemia virus (M-MuLV). The 4 X 10(6)-dalton classof RNA from M-MSV (MuLV) was examined for base sequence homology with DNA complementary to the 7 X 10(6)-dalton M-MuLV RNA genome. Approximately 86% of the M-MSV (MuLV) was protected from RNase digestion by hybridization, whereas 95% of M-MuLV was protected under identical conditions. These results indicate that the small RNA class of high-MOI M-MSV (MuLV) contains little (perhaps 10%) genetic information not present in M-MuLV. Virtually all of the 1.8 X 10(6)-dalton subunits of M-MSV (MuLV) RNA contained regions of poly(A) since 94% of the RNA bound to oligo(dT) cellulose in 0.5 M KCl. This suggests that the formation of the 1.8 X 10(6)-dalton subunits occurs before their packaging...
Proceedings of the National Academy of Sciences, 1975
We have studied the nucleic acid sequences in nonproducer cells transformed by Moloney sarcoma virus or Abeson leukemia virus (two ts of replication-defective, RNA-containing, viruses isolated by passage of Moloney leukemia virus in BALB/c mice). DNA probes from the Moloney leukemia virus detect RNA in both Abelson virus-transformed nonproducer cells and Moloney sarcoma virus-transformed nonproducer cells. A sarcoma-specific cDNA, prepared from the Moloney sarcoma virus, has extensive homology to RNA found in heterologous nonproducer cells transformed by. Moloney sarcoma virus, has little homology to RNA in cells producing Moloney leukemia virus, and no detectable homology to RNA in nonproducer cells transformed by the Abelson virus. By analogy to earlier data on avian and mammalian sarcoma viruses, these results suggest that the Moloney sarcoma virus arose by recombination between a portion of the Moloney leukemia virus genome and additional sarcoma-specific information, and indicate that the expression of this information is not essential for Abelson virusmediated fibroblast transformation.
Heteroduplex analysis of the RNA of clone 3 Moloney murine sarcoma virus
Journal of Virology
Heteroduplex analysis of the RNA isolated from purified virions of clone 3 Moloney murine sarcoma virus (M-MSV) hybridized to cDNA's from Moloney murine leukemia virus (M-MLV) and clone 124 M-MSV shows that the main physical component of clone 3 RNA is missing all or most of the 1.5-kilobase (kb) clone 124 M-MSV specific sequence denoted 13s (S. Hu et al. Cell 10 :469-477, 1977). This sequence is either deleted in clone 3 RNA or substituted by a very short (0.3-kilobase) sequence. In other respects, clone 3 and clone 124 RNAs show the same heteroduplex structure relative to M-MLV. Since Ps is believed to contain the src gene(s) of clone 124 RNA, this result leaves as an unresolved question the nature of the src gene(s) of the clone 3 M-MSV RNA complex.
Journal of General Virology, 1983
Intracellular polyadenylated viral RNA from cells infected by five different isolates of Moloney murine sarcoma virus (Mo-MuSV) was analysed by gel transfer hybridization. Genomic sizes of 4-6 kilobases (kb) for the ml-MuSV isolate, 5-2 kb for the m3and 124-MuSV, 6.1 kb for the HT1-MuSV and 6-7 kb for the 78-A1-MuSV were determined. With the exception of the ml strain, subgenomic RNA species were found in cells infected by the various isolates. However, no common subgenomic RNA containing v-mos sequences could be characterized. Each transformed cell line expressed a different set of viral RNA species in terms of size and structure.
Proceedings of the National Academy of Sciences, 1979
The unintegrated circular DNA form of Moloney murine sarcoma virus (MSV) has been cloned in bacteriophage X. Discrete deletions in the viral genome were shown to occur during propagation of recombinant phage in Escherichia coli. Heteroduplex and restriction enzyme analyses indicated the deletion of tandemly repeated sequences within certain of the cloned MSV DNA inserts. Cloned MSV DNA was used to prepare a probe composed of its acquired cellular (src) sequences, shown previously to be necessary for MSV transformation. Analysis of EcoRI digests of normal mouse cellular DNA revealed the presence of a single 14-kilobase-pair fragment containing these sequences which lacked contiguity with endogenous type C helper viral information of the same cells.
Base sequence differences between the RNA components of Harvey sarcoma virus
Journal of Virology
The 50 to 70S RNA of the Harvey sarcoma-Moloney leukemia virus (MLV) complex consists of 30 to 40S RNA subunits of two different size classes and contains sequences homologous to Moloney mouse leukemia virus and to information contained in a C-type rat virus, termed NRK virus. We have isolated by preparative gel electrophoresis the large (component 1) and the small (component 2) 30 to 40S RNA species from the Harvey sarcoma-MLV complex. Harvey RNA component 1 was completely complementary to DNA transcribed from MLV RNA and showed no homology to DNA transcribed from NRK virus when annealed under conditions of DNA excess. Harvey RNA component 2 was about 65% complementary to MLV DNA and about 33% complementary to NRK virus DNA.
Journal of Virology
Kirsten murine sarcoma-leukemia virus (Ki-MSV [MLV]) was found to contain less RNase H per unit of viral DNA polymerase than avian Rous sarcoma virus (RSV). Upon purification by chromatography on Sephadex G-200 and subsequent glycerol gradient sedimentation the avian DNA polymerase was obtained in association with a constant amount of RNase H. By contrast, equally purified DNA polymerase of Ki-MSV(MLV) and Moloney [Mo-MSV(MLV)] lacked detectable RNase H if assayed with two homopolymer and phage fd DNA-RNA hybrids as substrates. On the basis of picomoles of nucleotides turned over, the ratio of RNase H to purified avian DNA polymerase was 1: 20 and that of RNase H to purified murine DNA polymerase ranged between <1:2,800 and 5,000. Based on the same activity with poly (A) oligo(dT) the activity of the murine DNA polymerase was 6 to 60 times lower than that of the avian enzyme with denatured salmon DNA template or with avian or murine viral RNA templates assayed under various conditions (native, heat-dissociated, with or without oligo(dT) and oligo(dC) and at different template enzyme ratios). The template activities of Ki-MSV(MLV) RNA and RSV RNA were enhanced uniformly by oligo(dT) but oligo(dC) was much less efficient in enhancing the activity of MSV(MLV) RNA than that of RSV RNA. It was concluded that the purified DNA polymerase of Ki-MSV(MLV) differs from that of Rous sarcoma virus in its lack of detectable RNase H and in its low capacity to transcribe viral RNA and denatured salmon DNA. Some aspects of these results are discussed.
Isolation of a transformation-defective deletion mutant of Moloney murine sarcoma virus
Journal of Virology, 1982
A transformation-defective (td) deletion mutant of Moloney murine sarcoma virus (td Mo-MSV) and a transforming component termed Mo-MSV 3 were cloned from a stock of clone 3 Mo-MSV. To define the defect of the transforming function, the RNA of td Mo-MSV was compared with those of Mo-MSV 3 and of another transforming variant termed Mo-MSV 124 and with helper Moloney murine leukemia virus (Mo-MuLV). The RNA monomers of td Mo-MSV and Mo-MSV 3 comigrated on polyacrylamide gels and were estimated to be 4.8 kilobases (kb) in length. In agreement with previous analyses, the RNA of Mo-MSV 124 measured 5.5 kb and that of Mo-MuLV measured 8.5 kb. The interrelationships among the viral RNAs were studied by fingerprinting and mapping of RNase Tl-resistant oligonucleotides (Tl-oligonucleotides) and by identification of T1-oligonucleotides present in hybrids formed by a given viral RNA with cDNA's made from another virus. The nontransforming td Mo-MSV RNA lacked most of the Mo-MSV-specific sequence, i.e., the four 3'-proximal T1oligonucleotides of the six T1-oligonucleotides that are shared by the Mo-MSVspecific sequences of Mo-MSV 3 and Mo-MSV 124. The remaining two Mo-MSVspecific oligonucleotides identified td Mo-MSV as a deletion mutant of MSV rather than a deletion mutant of Mo-MuLV. td Mo-MSV and Mo-MSV 124 exhibited similar deletions of gag, pol, and env sequences which were less extensive than those of Mo-MSV 3. Hence, td Mo-MSV is not simply a deletion mutant of Mo-MSV 3. In addition to their MSV-specific sequences, all three MSV variants, including td Mo-MSV, shared the terminal sequences probably encoding the proviral long terminal repeat, which differed from their counterpart in Mo-MuLV. This may indirectly contribute to the oncogenic potential of MSV. A comparison of td Mo-MSV sequences with either Mo-MSV 124 or Mo-MSV 3 indicated directly, in a fashion similar to the deletion analyses which defined the src gene of avian sarcoma viruses, that Mo-MuLV-unrelated sequences of Mo-MSV are necessary for transformation. A definition of transformation-specific sequences of Mo-MSV by deletion analysis confirmed and extended previous analyses which have identified Mo-MuLV-unrelated sequences in Mo-MSV RNA and other studies which have described transformation of mouse 3T3 fibroblasts upon transfection with DNAs containing the Mo-MSV-specific sequence.
Journal of Virology
The large RNase T,-resistant oligonucleotides of the nondefective (nd) Rous sarcoma viruses (RSV): Prague RSV of subgroup B (PR-B), PR-C and B77 of subgroup C; of their transformation-defective (td) deletion mutants: td PR-B, td PR-C, and td B77; and of replication-defective (rd) RSV( -) were completely or partially mapped on the 30 to 40S viral RNAs. The location of a given oligonucleotide relative to the poly(A) terminus of the viral RNAs was directly deduced from the smallest size of the poly(A)-tagged RNA fragment from which it could be isolated. Identification of distinct oligonucleotides was based on their location in the electrophoretic/chromatographic fingerprint pattern and on analysis of their RNase A-resistant fragments. The following results were obtained. (i) The number of large oligonucleotides per poly(A)-tagged fragment increased with increasing size of the fragment. This implies that the genetic map is linear and that a given RNase T1-resistant oligonucleotide has, relative to the poly(A) end, the same location on all 30 to 40S RNA subunits of a given 60 to 70S viral RNA complex. (ii) Three sarcoma-specific oligonucleotides were identified in the RNAs of PR-B, PR-C and B77 by comparison with the RNAs of the corresponding td viruses. The sarcoma-specific oligonucleotides of these three sarcoma viruses had very similar compositions. Based on the map positions of these oligonucleotides, sarcoma-specific sequences of nd viral RNAs were estimated to map between 6.6 and 20% away from the poly(A) end. (iii) As far as analyzed, the oligonucleotide maps of the RNAs of nd and td PR-B were the same with the exception of the sarcoma-specific sequences, as were the maps of nd and td B77 RNAs. (iv) Comparisons of the map locations of oligonucleotides from the three nd sarcoma virus strains analyzed suggested that homologous oligonucleotides are found in certain homologous map positions of the respective RNAs. This implies that these three virus strains probably have similar gene orders. (v) The complexity in daltons of the RNA of PR-C was 3.22 x 106 and that of B77 was 3.02 x 106. (vi) The RNase T,-resistant oligonucleotides of poly(A)-tagged RNA fragments ranging up to 15S from each nd/td virus pair studied here were very similar. This implies that they share a common heteropolymeric sequence at their poly(A) ends. (vii) Poly(A)-tagged 12S RNA fragments of all avian tumor viruses studied so far including that of RSV(-) shared one oligonucleotide, termed spot C, which mapped very near the poly(A) end. It may be part of a short terminal heteropolymeric sequence common to all avian tumor virus RNAs investigated. The following maps emerged for the poly(A) terminal sequences of the nd/td virus RNAs which were analyzed: they start with poly(A) (molecular weight = 60,000), continue with a heteropolymeric sequence shared partially or completely by nd and td viruses (molecular weight = 140,000), which is followed by sarcoma-specific sequences (molecular weight 300,000 to 450,000) in the case of nd sarcoma virus RNAs. Several mechanisms are discussed to explain the generation of deletion mutants from nd viruses. 1051