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 · Sun Feb 01 00:00:00 EST 1981 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.78.2.838· OSTI ID:6357142

Rapaport, E ^[1]; Zamecnik, P C; Baril, E F

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1. 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), Vol. 78:2
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
DNA
POLYMERASES
LIGASES
BIOCHEMICAL REACTION KINETICS
TRANSFER RNA
AMINO ACIDS
ATP
CHEMICAL ACTIVATION
CHEMICAL BONDS
CHROMATOGRAPHY
DNA REPLICATION
ELECTROPHORESIS
ENZYME ACTIVITY
EXPERIMENTAL DATA
FRACTIONATION
HELA CELLS
MOLECULAR WEIGHT
TRYPTOPHAN
AZOLES
CARBOXYLIC ACIDS
DATA
ENZYMES
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
INDOLES
INFORMATION
KINETICS
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
NUMERICAL DATA
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PYRROLES
REACTION KINETICS
RNA
SEPARATION PROCESSES
TRANSFERASES
550201* - Biochemistry- Tracer Techniques