Further characterization of intracellular precursor polyproteins of rauscher leukemia virus

Jamjoom, Ghazi A.; Naso, Robert; Arlinghaus, Ralph B.

doi:10.1016/0042-6822(77)90075-7

Cited by 164 publications

(125 citation statements)

References 36 publications

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“…These two proteins can account for all of the [35S]methionine-labeled peptides present in the polyprotein. Taken in conjunction with previously reported observations (1,2,(6)(7)(8)(9)(10), these results directly demonstrate the gene for AMV DNA polymerase is adjacent to the 5'-proximal gag gene. This defines the structural gene order within the viral genome and suggests a scheme for the processing of the AMV DNA polymerase in vivo.…”

supporting

confidence: 79%

“…The structural proteins of the retraviruses are synthesized in vivo as large polypeptide precursors which are cleaved subsequently by proteolysis to the structural proteins characteristic of the mature virus particle (1,2). Similarly, we, as well as others, have demonstrated that the f subunit of the avian myeloblastosis virus (AMV) DNA polymerase (DNA nucleotidyltransferase, EC 2.7.7.7) is the precursor to the a subunit and that the a subunit is derived proteolytically from the fi subunit (3)(4)(5).…”

mentioning

confidence: 99%

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Cell-free synthesis of the precursor polypeptide for avian myeloblastosis virus DNA polymerase.

Paterson¹,

Marciani²,

Papas³

1977

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

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ABSTRACIHigh molecular weight RNA (35S) isolated from avian myeloblastosis virus directs the cell-free synthesis of two prominent polypeptides of 180,000 and 76,000 molecular weight. The latter polypeptide has previously been identified as the precursor to the group-specific antigens of the virus ("gag" proteins) [Vogt, V. M., Eisenman, R. & Diggelmann, H. (1975) J. Mol. Biol. 96,. Two-dimensional tryptic peptide analyses of the [15S]methionine-labeled peptides demonstrate that the 180,000-dalton product is a polyprotein that can account for all the peptides of the avian myeloblastosis virus DNA polymerase (DNA nucleotidyltransferase, EC 2.7.7.7.) and those of the gag viral proteins. This is direct confirmation of the genomic order of the viral structural genes, placing the polymerase gene adjacent to the 5'-proximal gag gene of the virus. Furthermore, our findings suggest that the primary polymerase gene product is the 3 subunit of the enzyme. These results are discussed in relation to the proposed structural gene map for the avian retraviruses and suggest a model for the in vivo processing of the viral polymerase.The structural proteins of the retraviruses are synthesized in vivo as large polypeptide precursors which are cleaved subsequently by proteolysis to the structural proteins characteristic of the mature virus particle (1, 2). Similarly, we, as well as others, have demonstrated that the f subunit of the avian myeloblastosis virus (AMV) DNA polymerase (DNA nucleotidyltransferase, EC 2.7.7.7) is the precursor to the a subunit and that the a subunit is derived proteolytically from the fi subunit (3-5). In the present communication we demonstrate that the ,B subunit of AMV DNA polymerase and the group-specific antigen ("gag") structural proteins of the virus are synthesized as a 180,000 molecular weight (Mr) polyprotein in a cell-free translational system. These two proteins can account for all of the [35S]methionine-labeled peptides present in the polyprotein. Taken in conjunction with previously reported observations (1, 2, 6-10), these results directly demonstrate the gene for AMV DNA polymerase is adjacent to the 5'-proximal gag gene. This defines the structural gene order within the viral genome and suggests a scheme for the processing of the AMV DNA polymerase in vivo.MATERIALS AND METHODS Virus Preparations. AMV was obtained from the plasma of infected chicks provided through contract number N01CP33291 under the Virus Cancer Program of the National Cancer Institute. The virus was concentrated and purified as described previously (11).Extraction of Viral RNA. Viral RNA was prepared from fresh viral pellets and extracted as described previously (12). Viral RNA was fractionated on sucrose gradients and the 70SThe costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertusemnent" in accordance with 18 U. S. C. §1734 solely to indicate this fact. 4951RNA fraction was precipitated with ethanol; pellets were dissolv...

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supporting

confidence: 79%

mentioning

confidence: 99%

Cell-free synthesis of the precursor polypeptide for avian myeloblastosis virus DNA polymerase.

Paterson¹,

Marciani²,

Papas³

1977

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

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“…On the basis of indirect evidence (12,13) and by analogy to avian type C viruses (14), the order of these genes within the mammalian type C viral genome appears to be 5'-gag-pol-env-3'. The primary translational product of the gag gene has been shown to be a 65,000-molecular weight (Mr) precursor that, upon posttranslational processing, gives rise to structural proteins of 30,000-(p30), 15,000-(pl5), 12,000-(p12), and 10,000-(plO) Mr (15,16). The internal arrangement of the gag gene has been…”

Section: Supeinfection Of Mink Cells Nonproductively Transformed Bymentioning

confidence: 99%

Cells nonproductively transformed by Abelson murine leukemia virus express a high molecular weight polyprotein containing structural and nonstructural components.

Reynolds¹,

Sacks²,

Deobagkar³

et al. 1978

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

114

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Cell clones nonproductively transformed by the replication-defective Abelson strain of murine leukemia virus (AbLV) were analyzed for type C viral antigen expression by competition immunoassay. AbLV-transformed mink non-producer lines were found to express a 110,000- to 130,000-molecular weight polyprotein containing murine leukemia virus gag proteins p15 and p12 covalently linked to nonstructural AbLV-coded component(s) of around 80,000-100,000 molecular weight. This polyprotein lacked detectable antigenic cross-reactivity with other virion-coded gag gene proteins such as p30, p10, the viral reverse transcriptase (RNA-dependent DNA polymerase), or the major viral envelope glycoprotein, gp70. By analogy to earlier data on feline and avian sarcoma viruses, these results suggest that a portion of this polyprotein might represent the AbLV src gene product and that in translation it is initially linked in precursor form to gag structural proteins. Superinfection of mink cells nonproductively transformed by AbLV--with either a wild mouse amphotropic type C virus isolate, 4070-A, or with the endogenous cat virus, RD114--led to production of pseudotype virus containing high concentrations of the AbLV-coded precursor polyprotein.

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“…An initiation site near the 5' end allows translation of the gag gene resulting in the synthesis of a proximal precursor protein (Pr65~°g), which upon post-translational cleavage gives rise to the structural proteins of the virion core, p30 the major component, p 15, p 12 and p 10 (Van Zaane et al, 1975 ;Jamjoom et al, 1977). The arrangement of the final virion proteins in Pr65 q°~ has ben established as N-p15-ppl2-p30-pl0 C (Oroszlan & Gilden, 1980).…”

Section: Introductionmentioning

confidence: 99%

Monoclonal Antibodies to Murine Retrovirus Protein p30

Chuat

Grce

Pilon

et al. 1985

Journal of General Virology

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SUMMARYHybrid cell lines were prepared by the fusion of mouse myeloma cells with the spleen cells of Wistar-Furth rats that had been immunized with a Moloney sarcoma virus (MoMuSV)-induced tumour, MFU. Two immunization protocols were designed. In the first, the animals received several injections of irradiated (10 000 rad) cells of a tumour cell line established in vitro, MFU-67. The rats received a booster injection 3 days prior to fusion. In the second protocol, immunization was the result of simple tumour growth, and no booster was given. Hybrids were tested by immunofluorescence for the production of immunoglobulins reacting with mouse cells acutely infected with MoMuSV. Over 20 ~ of reactive hybrids were observed in the tumour growth protocol, and about 10 ~o in the irradiated cell protocol when the last injection of the series was given 2 weeks before fusion. After 6 months, the proportion fell to 3~. Hybrid lines producing antibody to p30, the major core polypeptide of murine retroviruses, were obtained by cloning. Three of these were selected for closer study and were found to recognize three non-overlapping epitopes on p30. By direct and competitive binding in ELISA tests, the three epitopes were found to have very different distribution patterns among the various strains and isolates of murine retroviruses.

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Further characterization of intracellular precursor polyproteins of rauscher leukemia virus

Cited by 164 publications

References 36 publications

Cell-free synthesis of the precursor polypeptide for avian myeloblastosis virus DNA polymerase.

Cell-free synthesis of the precursor polypeptide for avian myeloblastosis virus DNA polymerase.

Cells nonproductively transformed by Abelson murine leukemia virus express a high molecular weight polyprotein containing structural and nonstructural components.

Monoclonal Antibodies to Murine Retrovirus Protein p30

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