Bovine papillomavirus type 2 (BPV-2) E5 oncoprotein binds to the subunit D of the V1-ATPase proton pump in naturally occurring urothelial tumors of the urinary bladder of cattle

Sante Roperto, Valeria Russo, Giuseppe Borzacchiello, Chiara Urraro, Roberta Lucà, Iolanda Esposito, Marita Georgia Riccardi, Cinzia Raso, Marco Gaspari, Dora Maria Ceccarelli, Rocco Galasso, Franco Roperto

Research output: Contribution to journalArticle

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Abstract

Background: Active infection by bovine papillomavirus type 2 (BPV-2) was documented for fifteen urinary bladder tumors in cattle. Two were diagnosed as papillary urothelial neoplasm of low malignant potential (PUNLMP), nine as papillary and four as invasive urothelial cancers. Methods and Findings: In all cancer samples, PCR analysis revealed a BPV-2-specific 503 bp DNA fragment. E5 protein, the major oncoprotein of the virus, was shown both by immunoprecipitation and immunohistochemical analysis. E5 was found to bind to the activated (phosphorylated) form of the platelet derived growth factor β receptor. PDGFβR immunoprecipitation from bladder tumor samples and from normal bladder tissue used as control revealed a protein band which was present in the pull-down from bladder cancer samples only. The protein was identified with mass spectrometry as "V1-ATPase subunit D", a component of the central stalk of the V1-ATPase vacuolar pump. The subunit D was confirmed in this complex by coimmunoprecipitation investigations and it was found to colocalize with the receptor. The subunit D was also shown to be overexpressed by Western blot, RT-PCR and immunofluorescence analyses. Immunoprecipitation and immunofluorescence also revealed that E5 oncoprotein was bound to the subunit D. Conclusion: For the first time, a tri-component complex composed of E5/PDGFβR/subunit D has been documented in vivo. Previous in vitro studies have shown that the BPV-2 E5 oncoprotein binds to the proteolipid c ring of the V0-ATPase sector. We suggest that the E5/PDGFβR/subunit D complex may perturb proteostasis, organelle and cytosol homeostasis, which can result in altered protein degradation and in autophagic responses.

Original languageEnglish
Article numbere88860
JournalPLoS One
Volume9
Issue number2
DOIs
Publication statusPublished - Feb 24 2014

Fingerprint

Bovine papillomavirus 1
Proton Pumps
proton pump
Oncogene Proteins
bladder
Immunoprecipitation
Urinary Bladder Neoplasms
adenosinetriphosphatase
Adenosine Triphosphatases
Tumors
Urinary Bladder
fluorescent antibody technique
neoplasms
Fluorescent Antibody Technique
cattle
Vacuolar Proton-Translocating ATPases
Proteolipids
Platelet-Derived Growth Factor Receptors
Neoplasms
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Bovine papillomavirus type 2 (BPV-2) E5 oncoprotein binds to the subunit D of the V1-ATPase proton pump in naturally occurring urothelial tumors of the urinary bladder of cattle. / Roperto, Sante; Russo, Valeria; Borzacchiello, Giuseppe; Urraro, Chiara; Lucà, Roberta; Esposito, Iolanda; Riccardi, Marita Georgia; Raso, Cinzia; Gaspari, Marco; Ceccarelli, Dora Maria; Galasso, Rocco; Roperto, Franco.

In: PLoS One, Vol. 9, No. 2, e88860, 24.02.2014.

Research output: Contribution to journalArticle

Roperto, S, Russo, V, Borzacchiello, G, Urraro, C, Lucà, R, Esposito, I, Riccardi, MG, Raso, C, Gaspari, M, Ceccarelli, DM, Galasso, R & Roperto, F 2014, 'Bovine papillomavirus type 2 (BPV-2) E5 oncoprotein binds to the subunit D of the V1-ATPase proton pump in naturally occurring urothelial tumors of the urinary bladder of cattle', PLoS One, vol. 9, no. 2, e88860. https://doi.org/10.1371/journal.pone.0088860
Roperto, Sante ; Russo, Valeria ; Borzacchiello, Giuseppe ; Urraro, Chiara ; Lucà, Roberta ; Esposito, Iolanda ; Riccardi, Marita Georgia ; Raso, Cinzia ; Gaspari, Marco ; Ceccarelli, Dora Maria ; Galasso, Rocco ; Roperto, Franco. / Bovine papillomavirus type 2 (BPV-2) E5 oncoprotein binds to the subunit D of the V1-ATPase proton pump in naturally occurring urothelial tumors of the urinary bladder of cattle. In: PLoS One. 2014 ; Vol. 9, No. 2.
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AU - Roperto, Sante

AU - Russo, Valeria

AU - Borzacchiello, Giuseppe

AU - Urraro, Chiara

AU - Lucà, Roberta

AU - Esposito, Iolanda

AU - Riccardi, Marita Georgia

AU - Raso, Cinzia

AU - Gaspari, Marco

AU - Ceccarelli, Dora Maria

AU - Galasso, Rocco

AU - Roperto, Franco

PY - 2014/2/24

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N2 - Background: Active infection by bovine papillomavirus type 2 (BPV-2) was documented for fifteen urinary bladder tumors in cattle. Two were diagnosed as papillary urothelial neoplasm of low malignant potential (PUNLMP), nine as papillary and four as invasive urothelial cancers. Methods and Findings: In all cancer samples, PCR analysis revealed a BPV-2-specific 503 bp DNA fragment. E5 protein, the major oncoprotein of the virus, was shown both by immunoprecipitation and immunohistochemical analysis. E5 was found to bind to the activated (phosphorylated) form of the platelet derived growth factor β receptor. PDGFβR immunoprecipitation from bladder tumor samples and from normal bladder tissue used as control revealed a protein band which was present in the pull-down from bladder cancer samples only. The protein was identified with mass spectrometry as "V1-ATPase subunit D", a component of the central stalk of the V1-ATPase vacuolar pump. The subunit D was confirmed in this complex by coimmunoprecipitation investigations and it was found to colocalize with the receptor. The subunit D was also shown to be overexpressed by Western blot, RT-PCR and immunofluorescence analyses. Immunoprecipitation and immunofluorescence also revealed that E5 oncoprotein was bound to the subunit D. Conclusion: For the first time, a tri-component complex composed of E5/PDGFβR/subunit D has been documented in vivo. Previous in vitro studies have shown that the BPV-2 E5 oncoprotein binds to the proteolipid c ring of the V0-ATPase sector. We suggest that the E5/PDGFβR/subunit D complex may perturb proteostasis, organelle and cytosol homeostasis, which can result in altered protein degradation and in autophagic responses.

AB - Background: Active infection by bovine papillomavirus type 2 (BPV-2) was documented for fifteen urinary bladder tumors in cattle. Two were diagnosed as papillary urothelial neoplasm of low malignant potential (PUNLMP), nine as papillary and four as invasive urothelial cancers. Methods and Findings: In all cancer samples, PCR analysis revealed a BPV-2-specific 503 bp DNA fragment. E5 protein, the major oncoprotein of the virus, was shown both by immunoprecipitation and immunohistochemical analysis. E5 was found to bind to the activated (phosphorylated) form of the platelet derived growth factor β receptor. PDGFβR immunoprecipitation from bladder tumor samples and from normal bladder tissue used as control revealed a protein band which was present in the pull-down from bladder cancer samples only. The protein was identified with mass spectrometry as "V1-ATPase subunit D", a component of the central stalk of the V1-ATPase vacuolar pump. The subunit D was confirmed in this complex by coimmunoprecipitation investigations and it was found to colocalize with the receptor. The subunit D was also shown to be overexpressed by Western blot, RT-PCR and immunofluorescence analyses. Immunoprecipitation and immunofluorescence also revealed that E5 oncoprotein was bound to the subunit D. Conclusion: For the first time, a tri-component complex composed of E5/PDGFβR/subunit D has been documented in vivo. Previous in vitro studies have shown that the BPV-2 E5 oncoprotein binds to the proteolipid c ring of the V0-ATPase sector. We suggest that the E5/PDGFβR/subunit D complex may perturb proteostasis, organelle and cytosol homeostasis, which can result in altered protein degradation and in autophagic responses.

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