Proteins encoded by the bovine papillomavirus E1 open reading frame: expression in heterologous systems and virally transformed cells

Sabine Santucci, Elliot Androphy, Catherine Bonne-Andréa, Philippe Clertant

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The E1 open reading frame (ORF) of bovine papillomavirus type 1 is required for the of viral genomes as multicopy plasmid molecules in transformed rodent firbroblasts. E1 has been reported to contain two seprate complementation groups (M and R, corresponding to N- C-terminal domains respectively) which regulate viral replication. However, E1 behaves as a single gene with respect to cell transformation and viral transcription. We examined the proteins translated from the entire ORF by using three antisera raised against E1 peptide or bacterial fusion proteins. The capacity of the whole ORF to encode a 72-kDa protein was demonstrated by translation of synthetic RNA in a reticulocyte lysate system, by microinjection of RNA into Xenopus oocytes, and by expression in recombinant baculoviruses and vaccinia viruses. In eucaryotic cells, this protine was fond to be phosphorylated and targeted to the cell nucleus. In vitro translation also produced shorter peptides, containing only the El C-terminal domain, because of internal translation starts on the third and fourth methionine codons within El ORF. On the other hand, mammalian cells infected by vaccinia E1 recombinant virus contained additional larger E1 phosphoproteins (transient 85-kDa stable 88-kDa species), likely representing processed forms of the 72-kDa species. The E1 72-kDa nuclear phosphoprotein was detected in bovine papillomavirus type 1-transformed cells. We report the biochemical characteristics of full-sized and truncated E1 proteins: (i) the C-terminal half of E1 ORF contains a phosphorylation site(s); (ii) the full-sized E1, but not the C-terminal protein, DNA, without for recognition of defined sequences, and critical determinants for this activity are likely confined to an N-terminal domain of the protein; (iii) covalent affinity labeling experiments performed on vaccinia virus-encoded E1 proteins with an ATP analog confirmed our previous observation of sequence similarities between the E1 C-terminal domain the ATPase of domain of simian virus 40 large T antigen.

Original languageEnglish (US)
Pages (from-to)6027-6039
Number of pages13
JournalJournal of Virology
Volume64
Issue number12
StatePublished - 1990
Externally publishedYes

Fingerprint

Bovine papillomavirus
Open Reading Frames
open reading frames
Bovine papillomavirus 1
Vaccinia virus
Phosphoproteins
Protein C
translation (genetics)
proteins
Viral Cell Transformation
cells
phosphoproteins
RNA
Vaccinia
Peptides
Proteins
Bacterial Proteins
Simian virus 40
Baculoviridae
Viral Genome

ASJC Scopus subject areas

  • Immunology

Cite this

Proteins encoded by the bovine papillomavirus E1 open reading frame : expression in heterologous systems and virally transformed cells. / Santucci, Sabine; Androphy, Elliot; Bonne-Andréa, Catherine; Clertant, Philippe.

In: Journal of Virology, Vol. 64, No. 12, 1990, p. 6027-6039.

Research output: Contribution to journalArticle

@article{e2e3944b63f14a4a9f235400b84c1057,
title = "Proteins encoded by the bovine papillomavirus E1 open reading frame: expression in heterologous systems and virally transformed cells",
abstract = "The E1 open reading frame (ORF) of bovine papillomavirus type 1 is required for the of viral genomes as multicopy plasmid molecules in transformed rodent firbroblasts. E1 has been reported to contain two seprate complementation groups (M and R, corresponding to N- C-terminal domains respectively) which regulate viral replication. However, E1 behaves as a single gene with respect to cell transformation and viral transcription. We examined the proteins translated from the entire ORF by using three antisera raised against E1 peptide or bacterial fusion proteins. The capacity of the whole ORF to encode a 72-kDa protein was demonstrated by translation of synthetic RNA in a reticulocyte lysate system, by microinjection of RNA into Xenopus oocytes, and by expression in recombinant baculoviruses and vaccinia viruses. In eucaryotic cells, this protine was fond to be phosphorylated and targeted to the cell nucleus. In vitro translation also produced shorter peptides, containing only the El C-terminal domain, because of internal translation starts on the third and fourth methionine codons within El ORF. On the other hand, mammalian cells infected by vaccinia E1 recombinant virus contained additional larger E1 phosphoproteins (transient 85-kDa stable 88-kDa species), likely representing processed forms of the 72-kDa species. The E1 72-kDa nuclear phosphoprotein was detected in bovine papillomavirus type 1-transformed cells. We report the biochemical characteristics of full-sized and truncated E1 proteins: (i) the C-terminal half of E1 ORF contains a phosphorylation site(s); (ii) the full-sized E1, but not the C-terminal protein, DNA, without for recognition of defined sequences, and critical determinants for this activity are likely confined to an N-terminal domain of the protein; (iii) covalent affinity labeling experiments performed on vaccinia virus-encoded E1 proteins with an ATP analog confirmed our previous observation of sequence similarities between the E1 C-terminal domain the ATPase of domain of simian virus 40 large T antigen.",
author = "Sabine Santucci and Elliot Androphy and Catherine Bonne-Andr{\'e}a and Philippe Clertant",
year = "1990",
language = "English (US)",
volume = "64",
pages = "6027--6039",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "12",

}

TY - JOUR

T1 - Proteins encoded by the bovine papillomavirus E1 open reading frame

T2 - expression in heterologous systems and virally transformed cells

AU - Santucci, Sabine

AU - Androphy, Elliot

AU - Bonne-Andréa, Catherine

AU - Clertant, Philippe

PY - 1990

Y1 - 1990

N2 - The E1 open reading frame (ORF) of bovine papillomavirus type 1 is required for the of viral genomes as multicopy plasmid molecules in transformed rodent firbroblasts. E1 has been reported to contain two seprate complementation groups (M and R, corresponding to N- C-terminal domains respectively) which regulate viral replication. However, E1 behaves as a single gene with respect to cell transformation and viral transcription. We examined the proteins translated from the entire ORF by using three antisera raised against E1 peptide or bacterial fusion proteins. The capacity of the whole ORF to encode a 72-kDa protein was demonstrated by translation of synthetic RNA in a reticulocyte lysate system, by microinjection of RNA into Xenopus oocytes, and by expression in recombinant baculoviruses and vaccinia viruses. In eucaryotic cells, this protine was fond to be phosphorylated and targeted to the cell nucleus. In vitro translation also produced shorter peptides, containing only the El C-terminal domain, because of internal translation starts on the third and fourth methionine codons within El ORF. On the other hand, mammalian cells infected by vaccinia E1 recombinant virus contained additional larger E1 phosphoproteins (transient 85-kDa stable 88-kDa species), likely representing processed forms of the 72-kDa species. The E1 72-kDa nuclear phosphoprotein was detected in bovine papillomavirus type 1-transformed cells. We report the biochemical characteristics of full-sized and truncated E1 proteins: (i) the C-terminal half of E1 ORF contains a phosphorylation site(s); (ii) the full-sized E1, but not the C-terminal protein, DNA, without for recognition of defined sequences, and critical determinants for this activity are likely confined to an N-terminal domain of the protein; (iii) covalent affinity labeling experiments performed on vaccinia virus-encoded E1 proteins with an ATP analog confirmed our previous observation of sequence similarities between the E1 C-terminal domain the ATPase of domain of simian virus 40 large T antigen.

AB - The E1 open reading frame (ORF) of bovine papillomavirus type 1 is required for the of viral genomes as multicopy plasmid molecules in transformed rodent firbroblasts. E1 has been reported to contain two seprate complementation groups (M and R, corresponding to N- C-terminal domains respectively) which regulate viral replication. However, E1 behaves as a single gene with respect to cell transformation and viral transcription. We examined the proteins translated from the entire ORF by using three antisera raised against E1 peptide or bacterial fusion proteins. The capacity of the whole ORF to encode a 72-kDa protein was demonstrated by translation of synthetic RNA in a reticulocyte lysate system, by microinjection of RNA into Xenopus oocytes, and by expression in recombinant baculoviruses and vaccinia viruses. In eucaryotic cells, this protine was fond to be phosphorylated and targeted to the cell nucleus. In vitro translation also produced shorter peptides, containing only the El C-terminal domain, because of internal translation starts on the third and fourth methionine codons within El ORF. On the other hand, mammalian cells infected by vaccinia E1 recombinant virus contained additional larger E1 phosphoproteins (transient 85-kDa stable 88-kDa species), likely representing processed forms of the 72-kDa species. The E1 72-kDa nuclear phosphoprotein was detected in bovine papillomavirus type 1-transformed cells. We report the biochemical characteristics of full-sized and truncated E1 proteins: (i) the C-terminal half of E1 ORF contains a phosphorylation site(s); (ii) the full-sized E1, but not the C-terminal protein, DNA, without for recognition of defined sequences, and critical determinants for this activity are likely confined to an N-terminal domain of the protein; (iii) covalent affinity labeling experiments performed on vaccinia virus-encoded E1 proteins with an ATP analog confirmed our previous observation of sequence similarities between the E1 C-terminal domain the ATPase of domain of simian virus 40 large T antigen.

UR - http://www.scopus.com/inward/record.url?scp=0025225428&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025225428&partnerID=8YFLogxK

M3 - Article

C2 - 2173778

AN - SCOPUS:0025225428

VL - 64

SP - 6027

EP - 6039

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 12

ER -