The low molecular weight RNAs of Rous sarcoma virus

Bishop, J. Michael; Levinson, Warren; Sullivan, Drew; Fanshier, Lois; Quintrell, Nancy; Jackson, Jeffrey J.

doi:10.1016/0042-6822(70)90251-5

Cited by 195 publications

(55 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the absence of this RNA primer molecule the template activity of RSV RNA is no better than that of other natural RNA molecules (14,17), suggesting that the preference exhibited by the RNA-directed DNA polymerase for oncornavirus 70S RNA as template is a special feature of the RNA and not a property of the enzyme. Although no virus-specific functions have previously been assigned to any of the other low molecular weight RNA species associated with virions of RNA tumor viruses, many of the virion-associated tRNA species appear to be cellular in origin and selectively incorporated into the virus particle during budding of the virus from the cell membrane (3,(7)(8)(9)(10)(11) (21). These results indicate that the 4S RNA primer molecule required by the RSV RNA-directed DNA polymerase for the initiation of DNA synthesis in vitro is present in normal, uninfected cells, and is in all likelihood a cell-specified molecule.…”

mentioning

confidence: 99%

“…The low-molecular-weight population includes 7S, 5S, and 4S RNA and is generally referred to as RSV-free (not associated with 70S RNA) RNA (1)(2)(3)(4)(5)(6). The free 4S RNA exhibits structural features of transfer RNA (tRNA) and can be acylated with amino acids (3,(5)(6)(7)(8)(9)(10)(11). RSV 70S RNA is segmented in nature, containing three to four high-molecular-weight 30-35S RNA subunits (1) and several low-molecular-weight species.…”

mentioning

confidence: 99%

See 1 more Smart Citation

RNA-directed DNA synthesis by the DNA polymerase of Rous sarcoma virus: structural and functional identification of 4S primer RNA in uninfected cells.

Faras¹,

Dibble²

1975

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

View full text Add to dashboard Cite

The RNA-directed DNA Virions of Rous sarcoma virus (RSV) contain two major size classes of RNA: 70S RNA presumed to be the viral genome, and a low-molecular-weight population consisting of a number of discrete RNA species. The low-molecular-weight population includes 7S, 5S, and 4S RNA and is generally referred to as RSV-free (not associated with 70S RNA) RNA (1-6). The free 4S RNA exhibits structural features of transfer RNA (tRNA) and can be acylated with amino acids (3, 5-11). RSV 70S RNA is segmented in nature, containing three to four high-molecular-weight 30-35S RNA subunits (1) and several low-molecular-weight species. This latter group includes 7S, 5S, and 4S RNA and is referred to as 70S-associated (70S-a) RNA (5,(12)(13)(14).Transcription in vitro of the 70S RNA genome of avian RNA tumor viruses by the RNA-directed DNA polymerase is initiated by covalent attachment of DNA to the 3' terminus of an RNA primer molecule (15-17). We have recently identified the RNA primer molecule as a low-molecularweight RNA species, 75 nucleotides in length, exhibiting structural features analogous to those of tRNA (18)(19)(20). This molecule appears to be the principal primer utilized by the RSV DNA polymerase, since its removal from 70S RNA results in the loss of 80-90% of the template activity of the 70S RNA complex (14, 17). In the absence of this RNA primer molecule the template activity of RSV RNA is no better than that of other natural RNA molecules (14, 17), suggesting that the preference exhibited by the RNA-directed DNA polymerase for oncornavirus 70S RNA as template is a special feature of the RNA and not a property of the enzyme. Although no virus-specific functions have previously been assigned to any of the other low molecular weight RNA species associated with virions of RNA tumor viruses, many of the virion-associated tRNA species appear to be cellular in origin and selectively incorporated into the virus particle during budding of the virus from the cell membrane (3, 7-11). It was of considerable interest, therefore, to determine whether the RNA primer molecule was a normal constituent of uninfected cells, or a virus-specified or virus-induced molecule.In

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

RNA-directed DNA synthesis by the DNA polymerase of Rous sarcoma virus: structural and functional identification of 4S primer RNA in uninfected cells.

Faras¹,

Dibble²

1975

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

View full text Add to dashboard Cite

show abstract

“…RNA was extracted from the infected ceils by the method of Bishop et al [9]: 15 ml of a solution con. taining 1% sodium dodecyl sulphate and 0.01 M EDTA in 0.05 M sodium acetate buffer (pH 5), was added to each bottle.…”

Section: Infection Of Cells and Purification Of Rnamentioning

confidence: 99%

Some characteristics of lower molecular weight arbovirus RNA's

et al. 1972

View full text Add to dashboard Cite

“…2, 10% polyacrylamide gel electrophoresis of the small RNA, both free and associated, yielded a number of bands (Fig. 2) representing different species of RNA, several of which have previously been identified [2,. One of the bands from the associated fraction, which had a mobility less than 5-S RNA, was particularly noticeable because of its relative intensity.…”

Section: Demonstration Of the Existence Of Viral 5943 R N Amentioning

confidence: 99%

“…It has been shown by separation on 10 acrylamide gel that the light fraction from sucrose gradients is heterogeneous and comprises transfer RNAs [l-31, 5-S RNA [4] and 8-S RNA [5,6]. It is generally accepted that all these RNAs are incorporated passively into the virions at the time of 'budding', and thus may have no special function.…”

mentioning

confidence: 99%

5.9‐S RNA, a New RNA Characterized in Several Mammalian Cell Lines

Galibert

Hampe

1977

European Journal of Biochemistry

View full text Add to dashboard Cite

A new species of RNA has been isolated from several different cell lines, both oncornavirus producing and non-producing. This RNA, which we designate 5.9-S RNA is present in the cellular cytoplasmic fraction at very low concentration (approximately 1 % of the quantity of 4-S RNA), but it accumulates to much higher levels in two murine oncornaviruses, Moloney murine sarcoma leukemia virus complex and Gross leukemia virus, where it represents as much as 10 % of the low-molecularweight RNA fraction associated with the 70-S RNA genome.The electrophoretic mobility and fingerprint analysis of T1 RNase digest products show that this species of RNA is approximately 160-165-residues long, and can be unequivocally distinguished from all previously described species of RNA in this size range.Analysis by sucrose gradient sedimentation of RNAs extracted from murine or avian oncornaviruses, has allowed the demonstration, in addition to 70-S RNA, of small RNAs and of ribosomal RNAs of cellular origin. It has been shown by separation on 10 acrylamide gel that the light fraction from sucrose gradients is heterogeneous and comprises transfer RNAs [l-31, 5-S RNA [4] and 8-S RNA [5,6]. It is generally accepted that all these RNAs are incorporated passively into the virions at the time of 'budding', and thus may have no special function. However, more recently, it has been shown that the synthesis of cDNA from 70-S RNA by reverse transcriptase uses an RNA primer which has been identified as tryptophan-tRNA in the avian system [7,8].In the course of an investigation designed to identify the RNA priming the transcription of the murine oncornavirus genome by reverse transcriptase, the presence in acrylamide gels of a band representing a new species of RNA was noted as a constant finding. This species of RNA, which we will refer to as 5.9-S RNA, is found in murine oncornaviruses, and also in cell lines, both virus-producing and non-producing. Its characterisation is the subject of the present report. MATERIALS AND METHODS MaterialsEnzymes and chemicals were obtained as described previously [9]. Wistar C F rat fibroblasts were prepared in Eagle's minimum essential medium and labelled with [32P]-orthophosphate during the first passage.Viruses were concentrated by ultracentrifugation of radioactive culture supernatants, and their RNA was extracted with phenol [12].Total cellular RNA was extracted with phenol as previously described 11 31. Subcellular fractions were prepared according to a technique modified from that of Penman [14]. The cells were suspended in buffer (0.01 M Tris, 0.01 M NaCI, 3 mM magnesium acetate, pH 7.4). After 30 min at 4 "C they were disrupted with a Dounce homogeniser and then centrifuged for 10 min at 10000 x g. The RNA of the 10000 x g supernatant was extracted after addition of 0.1 % sodium dodecyl sulphate and agitation with an equal volume of phenol. The 1OOOOxg pellet was suspended in buffer (0.01 M Tris, 0.5 M NaCI, 50 mM MgC12 pH 7.4) and 0.1 vol. of a sodium deoxycholate/ nonidet mixture ( 5 % sodium deoxyc...

show abstract

The low molecular weight RNAs of Rous sarcoma virus

Cited by 195 publications

References 24 publications

RNA-directed DNA synthesis by the DNA polymerase of Rous sarcoma virus: structural and functional identification of 4S primer RNA in uninfected cells.

RNA-directed DNA synthesis by the DNA polymerase of Rous sarcoma virus: structural and functional identification of 4S primer RNA in uninfected cells.

Some characteristics of lower molecular weight arbovirus RNA's

5.9‐S RNA, a New RNA Characterized in Several Mammalian Cell Lines

Contact Info

Product

Resources

About