Properties of a Ribonucleoprotein Particle Isolated from Nonidet P-40-Treated Rous Sarcoma Virus

Davis, Nancy; Rueckert, Roland R.

doi:10.1128/jvi.10.5.1010-1020.1972

Cited by 109 publications

(54 citation statements)

References 27 publications

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“…According to the current model for oncovirus structure, the location of the structural polypeptides of virions can be correlated with certain biochemical properties (6,33,34,45). To test this model with EIAV, the location of the virion polypeptides was determined by standard procedures in which purified virus was treated with protease (35) or detergent (14) and the resulting subparticles were isolated and analyzed for protein composition by SDS-PAGE.…”

Section: Resultsmentioning

confidence: 99%

“…Thus, gp90 and gp45 appear to form the surface projections of the virus, whereas p26, p15, pll, and p9 constitute the internal virion components. The identity of the internal protein(s) associated with the viral RNA was determined by treating purified EIAV with Nonidet P-40 to release the virion ribonucleoprotein complex, which was then isolated by centrifugation on a sucrose gradient (14). Typically, about 90% of the virion protein was solubilized by the detergent, and only 10% of the protein sedimented in the sucrose gradient in the position of the ribonucleoprotein subparticles.…”

Section: Resultsmentioning

confidence: 99%

“…The virion ribonucleoprotein complex was produced by treating purified EIAV with Nonidet P-40 (Shell Chemical Co., St. Louis, Mo. ), purified by centrifugation on a sucrose gradient, and analyzed for protein composition by SDS-PAGE (14).…”

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confidence: 99%

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Isolation and comparative biochemical properties of the major internal polypeptides of equine infectious anemia virus

Montelaro¹,

Lohrey²,

Parekh³

et al. 1982

J Virol

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We describe procedures for the large-scale production of equine infectious anemia virus (EIAV) and for the isolation of the four major non-glycosylated virion proteins, designated p26, p15, pll, and p9. Comparisons of the purified proteins by peptide mapping procedures and by enzyme-linked immunosorbent assays demonstrated the unrelatedness of the four proteins. The characteristic properties of each purified protein were examined by determining isoelectric points and amino acid compositions. We found that EIAV p26 and p9 focus at pl values of 6.2 and 5.0, respectively, and that these proteins contain no unusual amino acids. In contrast, EIAV p15 reproducibly displayed a heterogeneous isoelectric focusing pattern, with major pl values ranging from 5.7 to 8.3. This charge variation evidently correlated with different levels of phosphorylated serine or threonine or both, which could be detected by an amino acid analysis of purified p15. EIAV pll apparently focused at a pI of >10, reflecting its high content of basic amino acids. Moreover, localization experiments indicated that all four nonglycosylated proteins constitute the internal components of the virus, with all of the virion pll closely associated with the viral RNA genome. Thus, our results demonstrated that EIAV, a lentivirus, contains structural polypeptides which are analogous to the structural polypeptides described previously in prototype C oncoviruses.Equine infectious anemia is unique among virus-induced diseases in that clinical symptoms and associated bursts of viremia occur in sequential episodes separated by several weeks or months (13, 23). The available evidence indicates that the periodic nature of this disease is due to the sequential production and release of novel antigenic strains of equine infectious anemia virus (EIAV) which temporarily escape host immunosurveillance systems (13, 23-26). Thus, EIAV provides a dynamic system for investigating the interaction between host immune mechanisms and antigens involved in a persistent virus infection. However, a prerequisite for studying this system is a thorough description of the antigens of the causative virus.EIAV matures by budding from cytoplasmic membranes (31, 48), displays a complex morphology characteristic of type C viruses (16), and contains a reverse transcriptase (2) and a high-molecular-weight (60S to 70S) RNA genome composed of subunits (11). Based on these properties, EIAV has been tentatively classified as a member of the family Retroviridae (9, 13, 23, 46). However, serological comparisons have failed to detect any relatedness be-tween EIAV and a number of retroviruses (9, 46), and some ultrastructural studies (16) have suggested that EIAV resembles visna virus, a member of the lentivirus subfamily, more closely than any member of the oncovirus subfamily. These observations and the initial descriptions of EIAV and visna virus polypeptides have led to the suggestion that the protein composition of lentiviruses may be distinct from the protein compositions of prototype C mammalian ...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Isolation and comparative biochemical properties of the major internal polypeptides of equine infectious anemia virus

Montelaro¹,

Lohrey²,

Parekh³

et al. 1982

J Virol

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“…In this context, our understanding of the structure and functions of the retroviral nucleocapsid protein (i.e. NC) has come a long way since the first descriptions of oncoretroviruses and the oncoviral nucleoprotein (Ellermann and Bang, 1908;Rous, 1910;Davis and Rueckert, 1972). Initially studies on the retroviral NC focused on that of alpharetroviruses and gammaretroviruses, namely avian sarcoma leukosis viruses (ASLV) and the murine leukemia viruses (MuLV), respectively.…”

Section: Forewordsmentioning

confidence: 99%

“…The retroviral NC story began with the isolation of large ribonucleoprotein complexes (RNPs) (Davis and Rueckert, 1972) from purified viral particles of ASLV and MoMuLV, which were able to support viral DNA synthesis. These viral RNP complexes, consisting of the 60-70S genomic RNA dimer coated by about 2000 NC molecules, represent the most stable inner structure of the viral particle, with a circular chromatin-like conformation (Chen et al, 1980;Pager et al, 1994).…”

Section: Forewordsmentioning

confidence: 99%

Retrospective on the all-in-one retroviral nucleocapsid protein

et al. 2014

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This review aims at briefly presenting a retrospect on the retroviral nucleocapsid protein (NC), from an unspecific nucleic acid binding protein (NABP) to an all-in-one viral protein with multiple key functions in the early and late phases of the retrovirus replication cycle, notably reverse transcription of the genomic RNA and viral DNA integration into the host genome, and selection of the genomic RNA together with the initial steps of virus morphogenesis. In this context we will discuss the notion that NC protein has a flexible conformation and is thus a member of the growing family of intrinsically disordered proteins (IDPs) where disorder may account, at least in part, for its function as a nucleic acid (NA) chaperone and possibly as a protein chaperone vis-à-vis the viral DNA polymerase during reverse transcription. Lastly, we will briefly review the development of new anti-retroviral/AIDS compounds targeting HIV-1 NC because it represents an ideal target due to its multiple roles in the early and late phases of virus replication and its high degree of conservation.

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Small finger protein of avian and murine retroviruses has nucleic acid annealing activity and positions the replication primer tRNA onto genomic RNA.

et al. 1988

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Retrovirus virions carry a diploid genome associated with a large number of small viral finger protein molecules which are required for encapsidation. Our present results show that finger protein p12 of Rous sarcoma virus (RSV) and p10 of murine leukaemia virus (MuLV) positions replication primer tRNA on the replication initiation site (PBS) at the 5′ end of the RNA genome. An RSV mutant with a Val‐Pro insertion in the finger motif of p12 is able to partially encapsidate genomic RNA but is not infectious because mutated p12 is incapable of positioning the replication primer, tRNATrp. Since all known replication competent retroviruses, and the plant virus CaMV, code for finger proteins analogous to RSV p12 or MuLV p10, the initial stage of reverse transcription in avian, mammalian and human retroviruses and in CaMV is probably controlled in an analogous way.

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Properties of a Ribonucleoprotein Particle Isolated from Nonidet P-40-Treated Rous Sarcoma Virus

Abstract: medium containing 2%, fetal bovine serum and Dglucosamine-6-3H(N) (10 .Ci,/ml, 1 to 2 Ci 'mmole).

Cited by 109 publications

References 27 publications

Isolation and comparative biochemical properties of the major internal polypeptides of equine infectious anemia virus

Isolation and comparative biochemical properties of the major internal polypeptides of equine infectious anemia virus

Retrospective on the all-in-one retroviral nucleocapsid protein

Small finger protein of avian and murine retroviruses has nucleic acid annealing activity and positions the replication primer tRNA onto genomic RNA.

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