Expression of influenza hemagglutinin-polyoma T-antigen fusion proteins in a rat embryo fibroblast cell line

Kurt Ballmer-Hofer, Gail Mandel, Douglas V. Faller, Thomas M. Roberts, Thomas L. Benjamin

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Plasmids encoding the amino terminal portion of an influenza virus hemagglutinin (HA) fused to polyoma virus middle T (mT) or large T (lT) sequences have been constructed. Stable expression of the chimeric proteins was obtained in established rat embryo fibroblasts following plasmid co-transfection and selection for G418 resistance. The synthesis and localization of the proteins was followed by metabolic labeling with [35S]methionine and [3H]mannose, cell fractionation, and immunoprecipitation with anti-polyoma T antibody. The HA leader and amino terminal peptide direct the synthesis of the lT and mT proteins into the endoplasmic reticulum where they undergo glycosylation, but this occurs with a very low efficiency. Most of the HA-mT and HA-lT fusion protein molecules do not enter completely into the endoplasmic reticulum, but rather achieve their normal locations in the cell as slightly higher molecular weight proteins, presumably due to the extra sequences derived from HA at their amino termini. HA-mT fusion protein is found to have associated tyrosine-specific protein kinase activity precipitable with anti-src as well as anti-T antibody, and cells expressing this fusion protein have a transformed phenotype.

Original languageEnglish (US)
Pages (from-to)345-361
Number of pages17
JournalVirus Research
Volume6
Issue number4
DOIs
StatePublished - 1987
Externally publishedYes

Fingerprint

Viral Tumor Antigens
Hemagglutinins
Human Influenza
Embryonic Structures
Fibroblasts
Cell Line
Proteins
Endoplasmic Reticulum
Plasmids
Cell Fractionation
Polyomavirus
Cell Fusion
Mannose
Orthomyxoviridae
Glycosylation
Immunoprecipitation
Methionine
Protein-Tyrosine Kinases
Transfection
Anti-Idiotypic Antibodies

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Virology

Cite this

Expression of influenza hemagglutinin-polyoma T-antigen fusion proteins in a rat embryo fibroblast cell line. / Ballmer-Hofer, Kurt; Mandel, Gail; Faller, Douglas V.; Roberts, Thomas M.; Benjamin, Thomas L.

In: Virus Research, Vol. 6, No. 4, 1987, p. 345-361.

Research output: Contribution to journalArticle

Ballmer-Hofer, Kurt ; Mandel, Gail ; Faller, Douglas V. ; Roberts, Thomas M. ; Benjamin, Thomas L. / Expression of influenza hemagglutinin-polyoma T-antigen fusion proteins in a rat embryo fibroblast cell line. In: Virus Research. 1987 ; Vol. 6, No. 4. pp. 345-361.
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