PROCESSING AND STRUCTURE OF MURINE LEUKEMIA VIRUS gag AND env GENE ENCODED POLYPROTEINS

Oroszlan, Stephen; Henderson, Louis E.; Copeland, Terry D.; Schultz, Alan M.; Rabin, Evelyn

doi:10.1016/b978-0-12-417560-0.50023-4

Cited by 16 publications

(12 citation statements)

References 25 publications

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“…The very low level of these charged residues in the COOH-terminal 140 amino acids results in the moderate hydrophobicity of the chain in this region, as shown in Fig. 3B (26,27). REV-A also has a Thr-Ala doublet at this position (arrow …”

Section: Resultsmentioning

confidence: 98%

“…That this ATG is REV-A's env initiator is revealed by inspection of the REV-A DNA sequence and comparison with that of M-MuLV. In M-MuLV a 33-amino acid signal peptide containing a highly hydrophobic region is cleaved from the envelope precursor at a Thr-Ala linkage (26,27). In REV-A there is a Thr-Ala doublet at position [33][34], and the sequence between the methionine and this doublet contains 11 consecutive hydrophobic amino acids.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Nucleotide sequence of v-rel: the oncogene of reticuloendotheliosis virus.

Stephens¹,

Rice²,

Hiebsch³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

136

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The nucleotide sequence of v-rel, the oncogene carried by reticuloendotheliosis virus (REV), has been determined. The defective transforming genome REV arose through the insertion of v-rel (1,415 nucleotides) into the env gene of the helper virus REV-A. The predicted rel protein (503 amino acids) employs the REV-A enm initiator and terminates within the p2OE region of env. Because there are no natural antisera that detect the REV transforming protein, this nucleotide sequence provides the first step toward its isolation and characterization. The predicted protein is clearly distinct from all other reported transforming proteins but may be very distantly related to members of the arc family.mini by using [a-32P] Reticuloendotheliosis virus (REV), a type C retrovirus, causes acute leukemia in young chickens. The virus is a complex of a replication-competent helper virus called REV-A and a replication defective genome called REV, which is responsible for transformation (1). REV proviral DNA contains some sequences found in the helper virus, including long terminal repeats (LTRs) and parts of the gag, pol, and env genes, but it also contains an additional segment of about 1.5 kilobases (kb) not found in REV-A (2-6). This segment, termed v-rel, is the presumptive onc gene of the virus. Like the other known onc genes, v-rel appears to have been derived from a normal cellular gene, for related sequences have been found in DNA from uninfected avian species (5-9). How v-rel relates to the other onc genes has been unclear. No hybridization has been detected between REV and nucleic acids of other oncogenic viruses (refs. 5 and 8; M. Shibuya and H. Hanafusa, personal communication), and because to date there has been no antiserum that detects the v-rel protein (10, 11), its relatedness to other transforming proteins has been unknown. We report here the complete DNA sequence of v-rel. The protein predicted from this sequence is clearly distinct from all of the transforming proteins published to date but does appear to be very distantly related to members of the src family. MATERIALS AND METHODSThe cloning of Sac I fragments of REV and REV-A proviral DNA into AgtWES-AB has been described (6). For the purpose of sequence analysis various subclones of these DNA fragments were constructed in pBR322 or pUC8 (or both), both of which have been modified to include a Sac I site. A Charon 4A clone of REV-T (5) was a gift of I. Chen. Its internal 0.88-kb EcoRI fragment was subcloned into pBR322 and was used to confirm the sequence across the Sac I sites within v-rel.Restriction fragments of DNA were labeled at their 3' ter- RESULTS AND DISCUSSIONNucleotide Sequence of v-rel. The nucleotide sequence of v-rel and adjoining regions of REV-A-related proviral DNA is shown in Fig. 2 As shown in Fig. 3A, there is only one reading frame of vrel that is open for any significant length. This open frame begins slightly upstream from the 5' boundary of v-rel and extends through the entire transforming region to a termination codon (TGA)...

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Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Nucleotide sequence of v-rel: the oncogene of reticuloendotheliosis virus.

Stephens¹,

Rice²,

Hiebsch³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

136

View full text Add to dashboard Cite

show abstract

“…they are replication-competent, do not cause transformation of tissue culture cells, and both induce leukaemia in newborn mice after a prolonged period of latency, they do also have differences, i.e. their gp70 sequences differ (Oroszlan et al, 1980) and they induce tumours in different subsets of mouse lymphoid cells (Reddy et aL, 1980). Thus, the finding that there are different pIs for these two MuLV p30s should be considered along with known differences in gp70 and possible variability in the 5' or 3' long terminal repeated sequences when one is trying to understand how they induce different tumours.…”

Section: Discussionmentioning

confidence: 99%

Murine Leukaemia Virus p30 Heterogeneity as Revealed by Two-dimensional Gel Electrophoresis Is Not an Artefact of the Technique

Katoh

Yoshinaka

Luftig

1984

Journal of General Virology

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SUMMARYWe have utilized two-dimensional (2D) gel electrophoresis [the first dimension being a linear pH gradient (5 to 8) and the second an 8 to 15% acrylamide gradient] to characterize the virion protein, p30, from several strains of purified murine leukaemia virus (MuLV). In all cases, we found that there was a predominant (70 to 90%) Coomassie Brilliant Blue-staining p30 spot, as well as several other species which differed in pI. The major p30 spot differed in pI among different MuLV strains and the minor spots varied depending on the host cell used to grow the virus. Specifically, (i) Moloney (M)-MuLV/NIH-3T3 showed two spots, a major one at pI 6.3 and a more acidic one, (ii) AKR/NIH-3T3, AKR/mouse embryo, and Gross/NIH-3T3 showed four spots, with the two basic, minor spots of AKR/NIH-3T3 appearing relatively decreased in intensity, and (iii) Rauscher (R)-MuLV/JLS-V9 (BALB/c) showed two spots, a major one with greater than 90% of the estimated Coomassie Brilliant Blue stain at a pI of 6.5 and a minor, acidic one. The major spots of AKR and M-MuLV viruses also differed in pI. The major spot of the AKR and Gross N-tropic viruses had a pI of 6.7 while that of NB-tropic virus M-MuLV had a pI of 6.3. The possibility that the heterogeneity observed in p30 was an artefact of the 2D gel technique had to be considered since urea was used to denature proteins in the first dimension of the gel. This possibility was made unlikely by our finding that another technique, chromatofocusing, gave the same results. Specifically, M-MuLV/JLS-V9 p30, when separated on chromatofocusing columns under non-denaturing conditions yielded three peaks, each of which directly corresponded to the three spots (pI : 6.1, 6-3, 6.6) observed on 2D gels. Furthermore, tryptic peptide maps of the major (pI 6-3) and one of the minor (pI 6-6) M-MuLV spots, although very similar in peptide composition, showed about five clearly defined differences. These results indicate (i) that the p30s of several N-and NB-tropic viruses are heterogeneous in pI, and (ii) for one particular MuLV, the p30 heterogeneity can be explained by a difference in amino acid composition. These findings of p30 charge heterogeneity may reflect either the presence of several different p30s in each virus particle and/or a heterogeneity in the virus population.

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“…Although proteolytic activation of the fusion capacity has not been definitely shown before with FPV, this observation does not come as a surprise with the large body of information on other influenza hemagglutinins available. It should be pointed out, however, that there are numerous other viral glycoproteins, notably those of retroviruses, that undergo proteolytic processing at FPV hemagglutinin-like cleavage sites containing at least two basic amino acids (Oroszlan et al, 1980;Hunter et al, 1983;Koch et al, 1983;Wunsch etal., 1983;Adachi et al, 1984 (Bablanian et al, 1966 To analyze hemolytic activity, preparations of purified virus containing -16 hemagglutinating units (HAU) on 0.5 ml saline were added to 0.5 ml ammonium acetate buffer (0.5 M, pH 5.6) and 1.5 ml of a 1%7. suspension of human erythrocytes in saline.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of proteolytic cleavage of the hemagglutinin of influenza virus by the calcium-specific ionophore A23187.

Klenk¹,

Garten²,

Rott³

1984

The EMBO Journal

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At calcium-specific ionophore A23187 concentrations of -0.25 tM [which still allow assembly and release of fowl plague virus (FPV) particles] post-translational proteolytic cleavage of the viral hemagglutinin precursor HA into the fragments HA1 and HA2 is inhibited. The resulting virus particles with uncleaved hemagglutinin, that cannot be obtained under normal conditions, provide a suitable substrate for in vitro assays of the protease sensitivity of the FPV hemagglutinin. Proteolytic activation is accomplished with trypsin. Treatment with cathepsin B at low pH yields aberrant cleavage products suggesting that the cellular cleavage enzyme is not of lysosomal origin. A protease that cleaves the FPV hemagglutinin in the correct place can be detected in lysates of MDBK cells. This enzyme is calcium dependent and has a neutral pH optimum.

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PROCESSING AND STRUCTURE OF MURINE LEUKEMIA VIRUS gag AND env GENE ENCODED POLYPROTEINS

Cited by 16 publications

References 25 publications

Nucleotide sequence of v-rel: the oncogene of reticuloendotheliosis virus.

Nucleotide sequence of v-rel: the oncogene of reticuloendotheliosis virus.

Murine Leukaemia Virus p30 Heterogeneity as Revealed by Two-dimensional Gel Electrophoresis Is Not an Artefact of the Technique

Inhibition of proteolytic cleavage of the hemagglutinin of influenza virus by the calcium-specific ionophore A23187.

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