Analysis of adenovirus transforming proteins from early regions 1A and 1B with antisera to inducible fusion antigens produced in Escherichia coli
Plasmid vectors were constructed which expressed three adenovirus tumor antigens fused to a portion of the trpE protein of Escherichia coli. Insertion of adenovirus type 2-DNA from early region 1A (E1A) into such a plasmid led to a fusion protein which contained the C-terminal 266 amino acids of the 289-amino acid protein encoded by the viral 13S mRNA. Similarly, insertion of adenovirus type 5 DNA corresponding to the E1B 55- and 21-kilodalton proteins led to production of fusion proteins containing amino acid sequences from these proteins. Specific antisera for the E1A 289- and closely related 243-amino acid proteins and the E1B 55- and 21-kilodalton proteins were obtained. These sera were used to immunoprecipitate the tumor antigens in cells infected with wild-type and various mutants of adenovirus or to analyze them by an immunoblotting procedure. Mutant E1A proteins in which the C-terminal 70 amino acids are deleted were phosphorylated to much lower extents than the wild-type E1A proteins. This indicates that the deleted region is important for the process of phosphorylation. The E1A proteins were extracted, sedimented in glycerol gradients, analyzed by immunoprecipitation, and found to sediment primarily as monomers.
- Research Organization:
- Univ. of California, Los Angeles
- OSTI ID:
- 5942928
- Journal Information:
- J. Virol.; (United States), Journal Name: J. Virol.; (United States) Vol. 49:1; ISSN JOVIA
- Country of Publication:
- United States
- Language:
- English
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550701 -- Microbiology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADENOVIRUS
AMINO ACIDS
ANIMALS
ANTIGENS
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
DNA SEQUENCING
DRUGS
ESCHERICHIA COLI
EVEN-ODD NUCLEI
HELA CELLS
IMMUNE REACTIONS
IMMUNE SERUMS
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
METHIONINE
MICROORGANISMS
NUCLEI
ODD-ODD NUCLEI
ONCOGENIC VIRUSES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
OXYGEN COMPOUNDS
PARASITES
PHOSPHATES
PHOSPHORUS 32
PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
PHOSPHORYLATION
PLASMIDS
PROTEINS
RABBITS
RADIOASSAY
RADIOISOTOPES
STRUCTURAL CHEMICAL ANALYSIS
SULFUR 35
SULFUR ISOTOPES
VERTEBRATES
VIRUSES