Inhibition of protein synthesis by an RNA fraction from myeloblastosis virus

Olson, Kenneth; Deeney, A.O'c.; Beaudreau, G. S.

doi:10.1016/0005-2787(68)90129-9

Cited by 12 publications

(3 citation statements)

References 9 publications

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“…Our results reinforce previous reports that RNA from mammalian and plant viruses can be translated in a cell-free extract from E. coli (25)(26)(27)(28)(29)(30)(31)(32)(33)(34), a result that strongly favors the universality of the genetic code. Such a close correspondence between the proteins synthesized in animal cells and those synthesized in an E. coli lysate must imply that the sense codons of the two systems are largely identical.…”

Section: Analysis Of the Product By Immunodiffusionsupporting

confidence: 81%

Translation of Avian Myeloblastosis Virus RNA in a Cell-Free Lysate of Escherichia coli

Siegert

Konings

Bauer

et al. 1972

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

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In a cell-free extract of E. coli, RNA from avian myeloblastosis virus directs the synthesis of a protein that is antigenically identical with the group-specific antigen 4, and other proteins, three of which correspond in molecular weight to group-specific antigens 1-3. According to Temin's provirus hypothesis, the genetic information present in oncogenic RNA viruses is transcribed via an RNA-DNA hybrid into a double-stranded DNA molecule. After being incorporated into the host-cell genome, the DNA functions as a template for the synthesis of progeny RNA molecules (1). Several recent findings, such as the detection of reverse transcriptase (2, 3), host-controlled expression of virus genes (4, 5), rescue of latent viruses (6, 7), and vertical transmission of oncogenic viruses (8), strongly support this hypothesis.Apart from its function as a template for the synthesis of DNA, no clear role has been ascribed to the viral RNA; it is not known whether the information for the synthesis of virusspecific proteins is conserved in the viral RNA or in its complementary strand, or even in both.A hint that the viral RNA might have messenger properties is provided by the failure to detect complementary strands in the cytoplasm of infected cells. Complementary strands were detected only in the nucleus (9), and an RNA-dependent RNA polymerase was described (10). Whether or not a given RNA has messenger functions can be demonstrated more directly by a study of its ability to direct the synthesis of specific proteins in vitro. That this is the case for avian myeloblastosis virus (AMV)-RNA will be demonstrated in this paper by use of a cell-free lysate of Escherichia coli. MATERIALS AND METHODSPlasma-AMV, which was a generous gift of Dr. J. W. Beard (Duke University, Durham, N.C., USA) was first purified in a discontinuous sucrose gradient (11) with STE buffer (0.1 M NaCl-0.01 M Trise HCI (pH 8.0)-i mM ethylenediamine tetraacetate (EDTA). Virus present in the 35% sucrose layer was collected, diluted by the addition of one volume of NaCl-Tris-EDTA buffer, and pelleted by centrifugation (Spinco SW 50. 1; 45,000 rpm; 20 min at 40).The pellet was resuspended in NaCl-Tris-EDTA buffer and used for the preparation of viral RNA and viral proteins. The viral proteins were prepared as recently described for RNA-phage M12 proteins (12).RNA was extracted with a mixture of phenol and m-cresol (13) after incubation of the virus with Pronase in the presence of sodium dodecyl sulfate (SDS) at 370, according to a standard procedure (14). RNA present in the aqueous layer was precipitated twice with cold ethanol; high molecular weight RNA was obtained by linear sucrose density gradient centrifugation (15).The preparation of the cell-free lysate and of the incubation mixtures, and the extraction of the proteins synthesized in vitro, were already described (16).'4C-labeled proteins used in immunodiffusion tests in agar gel were prepared in the same manner (16), except that they were dialyzed against SSC (0.15 M NaCl-15 mM Nar citrate, pH 7...

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Section: Analysis Of the Product By Immunodiffusionsupporting

confidence: 81%

Translation of Avian Myeloblastosis Virus RNA in a Cell-Free Lysate of Escherichia coli

Siegert

Konings

Bauer

et al. 1972

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

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“…Evidence for this assumption was derived from the fact that this low molecular weight RNA fraction also inhibited the translation of phage Ml2 RNA (cf. [26]). Denaturation of RLV-RNA consistently resulted in an additional stimulation of the amino acid incorporation, while denaturation of MTV-RNA was much less effective.…”

Section: Effect Of Viral Rna and Eukaryotic Mrna On Amino Acid Incorpmentioning

confidence: 99%

Translation of oncogenic viral RNA and eukaryotic messenger RNA in the E. coli cell‐free system

et al. 1972

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“…Probably the same material has the capacity to incorporate methyl groups from methionine (Erikson, 1969), a property particular to cellular RNA (Brown and Attardi, 1967). The total 4-5 s viral RNA component possesses a unique property in that it is capable of inhibiting protein synthesis in vitro (Riman et al, 1967;Olson et al, 1968).…”

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confidence: 99%