The protein pattern of PMF virus, a type-D retrovirus from malignant permanent human cell lines

Uckert, Wolfgang; Wunderlich, V.; Bender, E; Sydow, G; Bierwolf, D

doi:10.1007/bf01318019

Cited by 15 publications

(16 citation statements)

References 11 publications

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“…1 shows the separation of proteins of purified t y p e D retroviruses after electrophoresis in a 7.5 to 15 per cent SDS polyacrylamide gradient slab gel. The patterns are similar to previously published d a t a for these viruses (4,18,19,24,27). A l t h o u g h the smaller proteins of each virus are designated as p l0, p12 and p15, respectively, it can be seen t h a t the molecular weights of these proteins differ slightly.…”

Section: Immunoprecipitation O/p25 O / M P M V and Helavsupporting

confidence: 88%

“…Viruses were purified by a combination of velocity gradient and isopycnic gradient centrifugation as described previously (24,25).…”

Section: Purification O/ Virusesmentioning

confidence: 99%

“…Polyacrylamide gradient slab gels (7.5--15 per cent) were prepared as described (24). About 60 ~g of purified viruses were dissociated in the presence of 1 per cent SDS and 0.1 per cent 2-mercaptoethanol by heating at 60 ° C for 5 minutes.…”

Section: Sds-polyacrylamide Slab Gel Electrophoresismentioning

confidence: 99%

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Further evidence from tryptic peptide analysis for heterogeneity among type D retroviruses

Uckert¹,

Hertling²,

Denner³

1984

Archives of Virology

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Tryptic peptide analyses were performed on the major internal structural proteins of type D retroviruses isolated originally from subhuman primate species (MPMV, SMRV, LV) and from permanent human cell lines (HeLaV, HEp-2V, PMFV). The p25 peptide maps of MPMV, LV, PMFV and HEp-2V were very similar but showed a striking dissimilarity to the p25 map of HeLaV and the p35 map of SMRV. All type D viruses included in this study could be distinguished by peptide maps of at least two of their three low-molecular-weight proteins p10, p12 and p15. These studies further demonstrate the heterogeneity among type D retroviruses.

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Section: Immunoprecipitation O/p25 O / M P M V and Helavsupporting

confidence: 88%

“…Viruses were purified by a combination of velocity gradient and isopycnic gradient centrifugation as described previously (24,25).…”

Section: Purification O/ Virusesmentioning

confidence: 99%

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Further evidence from tryptic peptide analysis for heterogeneity among type D retroviruses

Uckert¹,

Hertling²,

Denner³

1984

Archives of Virology

Self Cite

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“…They were labelled with 14C amino acid mixture (2 t~Ci/ml, The Radiochemical Centre Amersham, specific radioactivity 58 Ci/atom carbon) and ~H glucosamine (10 txCi/ml, The Radiochemical Centre Amersham, specific radioactivity 2.8 Ci/mmol) as described (25). Radiolabelling with 3:p phosphoric acid was performed according to S~ et al (21).…”

Section: Radiolabetling Of Viral Polypeptidesmentioning

confidence: 99%

Biochemical and immunological characterization of structural proteins from retrovirus-D/New England and comparison to Mason-Pfizer monkey virus and permanent human fibroblast virus

Uckert¹,

Wunderlich²,

Fiebach³

et al. 1987

Archives of Virology

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Biochemical and immunological properties of retrovirus-D/New England (here referred as R-D/NE) recently isolated at the New England Regional Primate Research Center, Southborough, Ma, from a rhesus monkey with acquired immune deficiency syndrome were investigated and compared to the prototype type D retroviruses Mason-Pfizer monkey virus (MPMV) and permanent human fibroblast virus (PMFV) isolated from a breast carcinoma of a rhesus monkey and a continuous human cell line, respectively. The polypeptide composition of R-D/NE propagated in a human lymphoid B cell line (Raji cells) has been investigated using SDS-polyacrylamide gel electrophoresis. Staining with Coomassie blue and labelling with 14C amino acids revealed seven viral polypeptides with molecular weights of 4,000, 10,000, 12,000, 15,000, 18,000, 27,000, and 80,000 Da which were also shared by MPMV and PMFV. The 80,000 Da protein was shown to be a glycoprotein by incorporation of 3H glucosamine. The 18,000 Da protein was identified as a phosphoprotein of R-D/NE. p18 structural proteins of MPMV and PMFV represent phosphoproteins of their respective viruses as well. All three phosphorylated proteins contain O-phosphoserine as major phosphoamino acid. The comparison of tryptic peptide maps of the major internal structural proteins of R-D/NE, MPMV, and PMFV revealed a striking similarity among p 10/p 12 and p 15. proteins. A minor difference was detected among the tryptic peptide digests of p 4 and p 27 proteins. Antiserum against p 15 of MPMV showed a significantly weaker binding to R-D/NE than to MPMV and PMFV at high dilutions.

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“…As with SFV and SIV, rare cases of genuine infection of primate handlers with SRVs have been described (282). In addition, SRVs grow efficiently in many human cell lines, and a number of reports that claimed the isolation of new human retrovirus turned out to be cases of laboratory contamination with SRV (19,262,496).…”

Section: Primate Betaretrovirusesmentioning

confidence: 99%

Human RNA “Rumor” Viruses: the Search for Novel Human Retroviruses in Chronic Disease

Voisset

Weiss

Griffiths

2008

Microbiol Mol Biol Rev

145

156

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SUMMARY Retroviruses are an important group of pathogens that cause a variety of diseases in humans and animals. Four human retroviruses are currently known, including human immunodeficiency virus type 1, which causes AIDS, and human T-lymphotropic virus type 1, which causes cancer and inflammatory disease. For many years, there have been sporadic reports of additional human retroviral infections, particularly in cancer and other chronic diseases. Unfortunately, many of these putative viruses remain unproven and controversial, and some retrovirologists have dismissed them as merely “human rumor viruses.” Work in this field was last reviewed in depth in 1984, and since then, the molecular techniques available for identifying and characterizing retroviruses have improved enormously in sensitivity. The advent of PCR in particular has dramatically enhanced our ability to detect novel viral sequences in human tissues. However, DNA amplification techniques have also increased the potential for false-positive detection due to contamination. In addition, the presence of many families of human endogenous retroviruses (HERVs) within our DNA can obstruct attempts to identify and validate novel human retroviruses. Here, we aim to bring together the data on “novel” retroviral infections in humans by critically examining the evidence for those putative viruses that have been linked with disease and the likelihood that they represent genuine human infections. We provide a background to the field and a discussion of potential confounding factors along with some technical guidelines. In addition, some of the difficulties associated with obtaining formal proof of causation for common or ubiquitous agents such as HERVs are discussed.

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The protein pattern of PMF virus, a type-D retrovirus from malignant permanent human cell lines

Cited by 15 publications

References 11 publications

Further evidence from tryptic peptide analysis for heterogeneity among type D retroviruses

Further evidence from tryptic peptide analysis for heterogeneity among type D retroviruses

Biochemical and immunological characterization of structural proteins from retrovirus-D/New England and comparison to Mason-Pfizer monkey virus and permanent human fibroblast virus

Human RNA “Rumor” Viruses: the Search for Novel Human Retroviruses in Chronic Disease

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