TY - JOUR
T1 - A 6700 MW membrane protein is encoded by region E3 of adenovirus type 2
AU - Wilson-Rawls, Jeanne
AU - Saha, Sankar K.
AU - Krajcsi, Peter
AU - Tollefson, Ann E.
AU - Gooding, Linda R.
AU - Wold, William S.M.
N1 - Funding Information:
We thank M. Pursley for technical asststance. This work was supported by Grants CA24710 (to W.S.M.W.), A126035, and CA48219 (to L.R.G. and W.S.M.W.) from the National Institutes of Health.
PY - 1990/9
Y1 - 1990/9
N2 - 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).
AB - 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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U2 - 10.1016/0042-6822(90)90395-8
DO - 10.1016/0042-6822(90)90395-8
M3 - Article
C2 - 1697127
AN - SCOPUS:0025071260
SN - 0042-6822
VL - 178
SP - 204
EP - 212
JO - Virology
JF - Virology
IS - 1
ER -