HeLa cell DNA polymerase. cap alpha. is tightly associated with tryptophanyl-tRNA synthetase and diadenosine 5',5'''-P/sup 1/,P/sup 4/-tetraphosphate binding activities
Journal Article
·
· Proc. Natl. Acad. Sci. U.S.A.; (United States)
- Boston Univ., MA
The purified high molecular weight form of HeLa cell DNA polymerase ..cap alpha.. (deoxynucleosidetriphosphate: DNA deoxynucleotidyltransferase, EC 2.7.7.7) was shown to associate tightly with several aminoacyl-tRNA synthetase activities. Fractionation of the high molecular weight enzyme on hexylagarose followed by gel filtration, chromatography on phosphocellulose, or polyacrylamide gel electrophoresis under nondenaturing conditions demonstrated copurification of only tryptophanyl-tRNA synthetase (L-tryptophan:tRNA/sup Trp/ ligase (AMP-forming), EC 6.1.1.2) along with DNA polymerase ..cap alpha... The high molecular weight (660,000) and low molecular weight (145,000) forms of DNA polymerase ..cap alpha.. were shown to possess a highly specific, noncovalent, diadenosine 5',5'''-P/sup 1/,P/sup 4/-tetraphosphate (Ap/sub 4/A) binding activity. The dissociation constants were determined to be 16 and 22 ..mu..M, respectively, by utilization of a charcoal adsorption procedure. No high-affinity binding of ATP could be detected. These findings suggest a link between the amino acid activation process and DNA replication in mammalian cells.
- OSTI ID:
- 6357142
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 78:2; ISSN PNASA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ATP
AZOLES
BIOCHEMICAL REACTION KINETICS
CARBOXYLIC ACIDS
CHEMICAL ACTIVATION
CHEMICAL BONDS
CHROMATOGRAPHY
DATA
DNA
DNA REPLICATION
ELECTROPHORESIS
ENZYME ACTIVITY
ENZYMES
EXPERIMENTAL DATA
FRACTIONATION
HELA CELLS
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
INDOLES
INFORMATION
KINETICS
LIGASES
MOLECULAR WEIGHT
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
NUMERICAL DATA
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PYRROLES
REACTION KINETICS
RNA
SEPARATION PROCESSES
TRANSFER RNA
TRANSFERASES
TRYPTOPHAN
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ATP
AZOLES
BIOCHEMICAL REACTION KINETICS
CARBOXYLIC ACIDS
CHEMICAL ACTIVATION
CHEMICAL BONDS
CHROMATOGRAPHY
DATA
DNA
DNA REPLICATION
ELECTROPHORESIS
ENZYME ACTIVITY
ENZYMES
EXPERIMENTAL DATA
FRACTIONATION
HELA CELLS
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
INDOLES
INFORMATION
KINETICS
LIGASES
MOLECULAR WEIGHT
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
NUMERICAL DATA
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PYRROLES
REACTION KINETICS
RNA
SEPARATION PROCESSES
TRANSFER RNA
TRANSFERASES
TRYPTOPHAN