Genetic mapping of a major site of phosphorylation in adenovirus type 2 E1A proteins
Adenovirus early region 1A (E1A) encodes two acidic phosphoproteins which are required for transactivation of viral transcription, efficient viral DNA replication in phase G/sub 0/-arrested human cells, and oncogenic transformation of rodent cells. Biochemical analysis of in vivo /sup 32/P-labeled adenovirus type 2 E1A proteins purified with monoclonal antibodies demonstrated that these proteins were phosphorylated at multiple serine residues. Two-dimensional phosphotryptic peptide maps of wild-type and mutant E1A proteins were used to locate a major site of E1A protein phosphorylation at serine-219 of the large E1A protein. Although this serine fell within a consensus sequence for phosphorylation by the cyclic AMP-dependent protein kinases, experiments with mutant CHO cells defective in these enzymes indicated that it was not. Oligonucleotide-directed mutagenesis was used to substitute an alanine for serine-219. This mutation prevented phosphorylation at this site. Nonetheless, the mutant was indistinguishable from the wild type for early gene transactivation, replication on G/sub 0/-arrested WI-38 cells, and transformation of cloned rat embryo fibroblast cells.
- Research Organization:
- Univ. of California, Los Angeles (USA)
- OSTI ID:
- 6046403
- Journal Information:
- J. Virol.; (United States), Journal Name: J. Virol.; (United States) Vol. 59:1; ISSN JOVIA
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
ADENOVIRUS
ALANINES
AMINO ACIDS
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CHO CELLS
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
ELECTROPHORESIS
EMBRYONIC CELLS
ENZYME ACTIVITY
FIBROBLASTS
GENETIC MAPPING
HYDROXY ACIDS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MAPPING
MICROORGANISMS
MONOCLONAL ANTIBODIES
MUTAGENESIS
NUCLEI
ODD-ODD NUCLEI
ONCOGENIC VIRUSES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PARASITES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORYLATION
PROTEINS
RADIOISOTOPES
RATS
RODENTS
SERINE
SOMATIC CELLS
TRANSCRIPTION
TWO-DIMENSIONAL ELECTROPHORESIS
VERTEBRATES
VIRUSES