A 6700 MW membrane protein is encoded by region E3 of adenovirus type 2

Norma Wilson-Rawls, Sankar K. Saha, Peter Krajcsi, Ann E. Tollefson, Linda R. Gooding, William S M Wold

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

There is an open reading frame between ATG1022 and TGA1205 in the E3 transcription unit of adenovirus 2 that could encode a protein of MW 6700 (6.7K) (61 amino acids). To address whether this protein is expressed, we prepared an antiserum against a synthetic peptide corresponding to residues 47-61 in the 6.7K protein. This antiserum immunoprecipitated two series of protein bands, a 7K-8K doublet and a 15K-16K doublet or triplet, as observed by electrophoresis on 10-18% gradient SDS-polyacrylamide gels. These bands were not obtained from cells infected with mutants that lack the 6.7K gene. Most, if not all, of the 7K-8K and 15K-16K bands were detected by immunoblot, indicating that they are modified versions of the 6.7K protein. Only an 8K band was observed after cell-free translation of hybridization-purified mRNA, suggesting that this may be the primary translation product. As judged by DNA sequence, the 6.7K protein has a hydrophobic domain of at least 22 residues (residues 16-37), suggesting that 6.7K may be a membrane protein. Consistent with this, the 7K-8K and 15K-16K bands were observed in the crude membrane but not the cytosol or nuclear fractions of biochemically fractionated cells. The 6.7K protein was underproduced by mutants which under-produce E3 mRNAs a and c, indicating that 6.7K is translated from these mRNAs. Since the E3-gp19K protein is also translated from mRNAs a and c, these mRNAs are bicistronic. The 6.7K protein is well-conserved in Ad5 (Ad2 and Ad5 are group C adenoviruses), and appears to be marginally conserved in Ad3 (group B).

Original languageEnglish (US)
Pages (from-to)204-212
Number of pages9
JournalVirology
Volume178
Issue number1
DOIs
StatePublished - 1990
Externally publishedYes

Fingerprint

Adenoviridae
Membrane Proteins
Proteins
Messenger RNA
Immune Sera
Cytosol
Open Reading Frames
Electrophoresis
Amino Acids
Peptides
Membranes
Genes

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Wilson-Rawls, N., Saha, S. K., Krajcsi, P., Tollefson, A. E., Gooding, L. R., & Wold, W. S. M. (1990). A 6700 MW membrane protein is encoded by region E3 of adenovirus type 2. Virology, 178(1), 204-212. https://doi.org/10.1016/0042-6822(90)90395-8

A 6700 MW membrane protein is encoded by region E3 of adenovirus type 2. / Wilson-Rawls, Norma; Saha, Sankar K.; Krajcsi, Peter; Tollefson, Ann E.; Gooding, Linda R.; Wold, William S M.

In: Virology, Vol. 178, No. 1, 1990, p. 204-212.

Research output: Contribution to journalArticle

Wilson-Rawls, N, Saha, SK, Krajcsi, P, Tollefson, AE, Gooding, LR & Wold, WSM 1990, 'A 6700 MW membrane protein is encoded by region E3 of adenovirus type 2', Virology, vol. 178, no. 1, pp. 204-212. https://doi.org/10.1016/0042-6822(90)90395-8
Wilson-Rawls, Norma ; Saha, Sankar K. ; Krajcsi, Peter ; Tollefson, Ann E. ; Gooding, Linda R. ; Wold, William S M. / A 6700 MW membrane protein is encoded by region E3 of adenovirus type 2. In: Virology. 1990 ; Vol. 178, No. 1. pp. 204-212.
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abstract = "There is an open reading frame between ATG1022 and TGA1205 in the E3 transcription unit of adenovirus 2 that could encode a protein of MW 6700 (6.7K) (61 amino acids). To address whether this protein is expressed, we prepared an antiserum against a synthetic peptide corresponding to residues 47-61 in the 6.7K protein. This antiserum immunoprecipitated two series of protein bands, a 7K-8K doublet and a 15K-16K doublet or triplet, as observed by electrophoresis on 10-18{\%} gradient SDS-polyacrylamide gels. These bands were not obtained from cells infected with mutants that lack the 6.7K gene. Most, if not all, of the 7K-8K and 15K-16K bands were detected by immunoblot, indicating that they are modified versions of the 6.7K protein. Only an 8K band was observed after cell-free translation of hybridization-purified mRNA, suggesting that this may be the primary translation product. As judged by DNA sequence, the 6.7K protein has a hydrophobic domain of at least 22 residues (residues 16-37), suggesting that 6.7K may be a membrane protein. Consistent with this, the 7K-8K and 15K-16K bands were observed in the crude membrane but not the cytosol or nuclear fractions of biochemically fractionated cells. The 6.7K protein was underproduced by mutants which under-produce E3 mRNAs a and c, indicating that 6.7K is translated from these mRNAs. Since the E3-gp19K protein is also translated from mRNAs a and c, these mRNAs are bicistronic. The 6.7K protein is well-conserved in Ad5 (Ad2 and Ad5 are group C adenoviruses), and appears to be marginally conserved in Ad3 (group B).",
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