Conservation of an ATP-binding domain among recA proteins from Proteus vulgaris, erwinia carotovora, Shigella flexneri, and Escherichia coli K-12 and B/r
The purified RecA proteins encoded by the cloned genes from Proteus vulgaris, Erwinia carotovora, Shigella flexneri, and Escherichia coli B/r were compared with the RecA protein from E. coli K-12. Each of the proteins hydrolyzed ATP in the presence of single-stranded DNA, and each was covalently modified with the photoaffinity ATP analog 8-azidoadenosine 5'-triphosphate (8N/sub 3/ATP). Two-dimensional tryptic maps of the four heterologous RecA proteins demonstrated considerable structural conservation among these bacterial genera. Moreover, when the (..cap alpha..-/sup 32/P)8N/sub 3/ATP-modified proteins were digested with trypsin and analyzed by high-performance liquid chromatography, a single peak of radioactivity was detected in each of the digests and these peptides eluted identically with the tryptic peptide T/sub 31/ of the E. coli K-12 RecA protein, which was the unique site of 8N/sub 3/ATP photolabeling. Each of the heterologous recA genes hybridized to oligonucleotide probes derived from the ATP-binding domain sequence of the E. coli K-12 gene. These last results demonstrate that the ATP-binding domain of the RecA protein has been strongly conserved for greater than 10/sup 7/ years.
- Research Organization:
- Massachusetts Institute of Technology, Cambridge (USA)
- OSTI ID:
- 6805011
- Journal Information:
- J. Bacteriol.; (United States), Vol. 170:6
- Country of Publication:
- United States
- Language:
- English
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PATHOGENS
BIOLOGICAL EVOLUTION
MOLECULAR BIOLOGY
PEPTIDE HYDROLASES
GENES
ATP
BIOCHEMICAL REACTION KINETICS
DNA-CLONING
GENETIC MAPPING
PHOSPHORUS 32
PROTEINS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CLONING
DAYS LIVING RADIOISOTOPES
ENZYMES
HYDROLASES
ISOTOPES
KINETICS
LIGHT NUCLEI
MAPPING
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
RADIOISOTOPES
REACTION KINETICS
550201* - Biochemistry- Tracer Techniques