Nucleotide sequence of the envelope gene of Friend murine leukemia virus

Koch, Werner; Hunsmann, Gerhard; Friedrich, Roland

doi:10.1128/jvi.45.1.1-9.1983

Cited by 138 publications

(72 citation statements)

References 58 publications

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“…reflecting its extremely hydrophobic properties. This assignment is corroborated by the observation that the corresponding peak fraction is also registered by monitoring the fluorescence due to the presence of two tryptophan residues in p12E [31] as well as by liquid scintillation counting (data not shown). Starting from cell culture supema- tants of 147 culture dishes, the average yield of purified p12E was about 44 pg.…”

Section: Resultssupporting

confidence: 59%

“…The deviation of 8 Da (0.04%) reflects a comparatively low mass accuracy, which might be due to adduct formation between the highly hydrophobic polypeptide and matrix or sample impurities [29]. The only N-terminal amino acid sequence determined was Glu-Pro-Val-Ser-Leu-Thr in agreement with the published N-terminus of pl2E [31]. Hence, it may be concluded that the purified protein represents pl2E.…”

Section: Resultssupporting

confidence: 53%

“…Since reducing agents such as 2-mercaptoethanol or dithiothreitol have been shown to release fatty acids from thioester linkages [32], SDS/PAGE was usually carried out under nonreducing conditions. [31] plus one covalently linked palmitic acid residue, an average molecular mass of 20056 Da can be calculated. The deviation of 8 Da (0.04%) reflects a comparatively low mass accuracy, which might be due to adduct formation between the highly hydrophobic polypeptide and matrix or sample impurities [29].…”

Section: Resultsmentioning

confidence: 99%

“…5). Amino acid sequence of p12E[31]. Tryptic cleavage sites are marked (4) ; resulting peptides are numbered.…”

mentioning

confidence: 99%

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Localization of the Palmitoylation Site in the Transmembrane Protein p12E of Friend Murine Leukaemia Virus

Hensel¹,

Hintz²,

Karas³

et al. 1995

Eur J Biochem

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Friend murine leukaemia virus complex was propagated on murine cells in the presence of 19,IO-'Hlpalmitic acid. Virus particles were harvested from the culture supernatant and lysed with detergents. The viral transmembrane protein, pl2E, was isolated from the lysates by size-exclusion chromatography and purified by narrowbore reverse-phase HPLC. Analysis of the purified product by matrix-assisted laser desorptiodionization time-of-flight mass spectrometry (MALDT-TOF-MS) revealed that the protein is palmitoylated carrying one fatty acid residue. The radiolabelled fatty acid was released by hydroxylamine treatment at pH 7, indicating that acylation occurred via a thioester linkage. For allocation of the acylation site, p12E was digested with trypsin. The resulting peptides were either directly subjected to MALDI-TOF-MS or fractionated by microbore reverse-phase HPLC prior to mass spectrometry. The results revealed that p12E of Friend murine leukaemia virus is acylated at a cysteine residue situated at the Cterminal side of the putative transmembrane anchor of the polypeptide. Fatty acid analysis of the purified acylpeptide demonstrated that p12E carries almost exclusively palmitic acid.Keywords: fatty acid; mass spectrometry ; protein acylation ; retrovirus.The envelope (env) gene products of Friend murine leukaemia virus (F-MuLV) are synthesized as a common precursor molecule, gPr90, that is proteolytically cleaved into the viral glycoprotein, gp71, and the nonglycosylated transmembrane protein, p15E [I]. In a second step, p15E is processed by a virusencoded protease yielding p12E [2, 31. The retroviral envelope glycoprotein mediates the first contact with specific host cell receptors [4-81 thus determining the viral host range 18, 91. The corresponding transmembrane protein, p12E, functions as an anchor for gp71. Furthermore, it is substantially involved in the fusion of viral and cellular membranes [lo, 111. It has been demonstrated that the membrane fusion capability is greatly enhanced by processing of p15E to p12E [12]. Hence, both env gene products play an important role in viral pathogenesis [13].Using metabolic labelling with ['Hlpalmitic acid, evidence has been provided that F-MuLV p12E might be post-translationally acylated [14]. However, there is no direct proof that a fatty acid is covalently linked to p12E. Furthermore, the fatty acid substitution pattern, the type of linkage, and the acylation site have not been investigated. Since palmitoylation of proteins has been reported to play an important role in controlling their intracellular location and biological functions [15 -181, we have analysed the acylation of F-MuLV p12E in detail. The results obCorrespondence to R. Geyer, Biochemisches Institut am Klinikum Fax: +49 641 702 4103. Abbreviations. CF,CO,H, trifluoroacetic acid ; F-MuLV, Friend murine leukaemia virus; gp71, 71-kDa glycoprotein; gPr90, primary translation product of the envelope gene; MALDI-TOF-MS, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry ; p12E and...

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Section: Resultssupporting

confidence: 59%

Section: Resultssupporting

confidence: 53%

Section: Resultsmentioning

confidence: 99%

“…5). Amino acid sequence of p12E[31]. Tryptic cleavage sites are marked (4) ; resulting peptides are numbered.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Localization of the Palmitoylation Site in the Transmembrane Protein p12E of Friend Murine Leukaemia Virus

Hensel¹,

Hintz²,

Karas³

et al. 1995

Eur J Biochem

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“…The biological activity of these MAbs was characterized by the use of two FLV clones with different tissue tropisms: the neurotropic A8 virus and the non-neurotropic FLV clone 57 virus (Oliff et al, 1980). The 57 virus was used as a non-neuropathogenic, erythroleukemogenic wild-type virus because its biological activity has been studied extensively (Masuda et al, 1992) and its nucleotide sequence is available (Koch et al, 1983).…”

mentioning

confidence: 99%

Characterization of monoclonal antibodies recognizing neurotropic Friend murine leukemia virus

1995

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We isolated a replication-competent, neurotropic retrovirus (FrC6 virus) and its molecular clone A8 from the NB-tropic Friend murine leukemia virus (FLV) complex. For detection and characterization of the FrC6 and A8 viruses, monoclonal antibodies (MAbs) against the FLV complex were established. Thirty MAbs, each of which reacted with the FLV-producing cell line, were tested for potential neutralizing activities; only two MAbs inhibited the proliferation of the A8 virus. These two MAbs were ineffective or had very weak neutralizing activities toward the non-neurotropic FLV strain clone 57 virus. Further characterization of MAbs by immunoprecipitation revealed that 4 MAbs recognized the envelope protein of the A8 virus. Two of these 4 MAbs recognized the surface glycoprotein gp70, requiring the conformational epitope of the virus for this recognition, while the other two MAbs, which were reactive with the transmembrane protein p15E, were conformation-independent. Both of the MAbs against gp70 distinguished neuropathogenic and non-neuropathogenic viruses to some extent, through neutralizing activity or binding activity detected by immunoprecipitation, whereas the two MAbs against p15E reacted with the viruses in a similar manner. Furthermore, one of the MAbs distinguished the viral antigen in the wall of the vacuolation that composes the spongiotic lesion induced by FrC6 viral infection of the brain.

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Sequence analysis of Gardner-Arnstein feline leukaemia virus envelope gene reveals common structural properties of mammalian retroviral envelope genes.

Wunsch¹,

Schulz²,

Koch³

et al. 1983

The EMBO Journal

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We have sequenced the envelope (env) gene and most of the adjacent 3′ long terminal repeat (LTR) of Gardner‐Arnstein feline leukaemia virus subtype B. The LTR of this virus contains, at corresponding positions, all signal sequence elements known from other retroviral LTRs. The deduced amino acid sequence of the longest open reading frame was compared with env polypeptide sequences of several murine leukaemia viruses. This allowed us to predict the positions of both p12/15env and gp70 polypeptides as well as a hydrophobic leader polypeptide. The env polypeptides of the different viruses show long stretches of homology and similar hydrophilicity profiles in the p12env region and in the carboxy‐terminal half of gp70 (constant region). The most extensive variations are confined to certain parts of the amino‐terminal half of gp70 (differential region). In this region, however, feline leukaemia virus and murine mink cell focus forming viruses are still closely related. A correspondingly spaced pattern of identical, short amino acid sequences appears in three different parts of the env polyprotein, suggesting its evolution from a primordial env‐related precursor by tandem duplications.

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Nucleotide sequence of the envelope gene of Friend murine leukemia virus

Cited by 138 publications

References 58 publications

Localization of the Palmitoylation Site in the Transmembrane Protein p12E of Friend Murine Leukaemia Virus

Localization of the Palmitoylation Site in the Transmembrane Protein p12E of Friend Murine Leukaemia Virus

Characterization of monoclonal antibodies recognizing neurotropic Friend murine leukemia virus

Sequence analysis of Gardner-Arnstein feline leukaemia virus envelope gene reveals common structural properties of mammalian retroviral envelope genes.

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