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Resolution of the diadenosine 5',5'''-P/sup 1/,P/sup 4/-tetraphosphate binding subunit from a multiprotein form of HeLa cell DNA polymerase. cap alpha

Journal Article · · Proc. Natl. Acad. Sci. U.S.A.; (United States)
; ; ; ;
A diadenosine 5',5'''-P/sup 1/,P/sup 4/-tetraphosphate (Ap/sub 4/A) binding subunit has been resolved from a high molecular weight (640,000) multiprotein form of DNA polymerase ..cap alpha.. (deoxy-nucleoside triphosphate:DNA nucleotidyltransferase (DNA-directed), EC 2.7.7.7) from HeLa cells. The Ap/sub 4/A binding activity copurifies with the DNA polymerizing activity during the course of purification. Hydrophobic chromatograpy on butylagarose resolves the Ap/sub 4/A binding activity from the DNA polymerase. The Ap/sub 4/A binding activity is protein in nature since the binding of Ap/sub 4/A is abolished by treatment of the isolated binding activity with proteinase K but is insensitive to treatment with DNase or RNase. The molecular weight of the Ap/sub 4/A binding protein, as determined by polyacrylamide gel electrophoresis under nondenaturing conditions or by NaDodSO/sub 4//polyacrylamide gel electrophoresis after photoaffinity labeling of the protein with (/sup 32/P)Ap/sub 4/A is 92,000 or 47,000. The binding activity of this protein is highly specific for Ap/sub 4/A.
Research Organization:
Worcester Foundation for Experimental Biology, Shrewsbury, MA
DOE Contract Number:
AC02-80EV10326
OSTI ID:
5932316
Journal Information:
Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 80; ISSN PNASA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHEMICAL BONDS
DATA
DAYS LIVING RADIOISOTOPES
DNA POLYMERASES
DNA REPLICATION
ENZYME ACTIVITY
ENZYMES
ESTERS
EXPERIMENTAL DATA
HELA CELLS
INFORMATION
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
MOLECULAR WEIGHT
NUCLEI
NUCLEIC ACID REPLICATION
NUCLEOTIDYLTRANSFERASES
NUMERICAL DATA
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PHOSPHORIC ACID ESTERS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PROTEINS
PURIFICATION
RADIOISOTOPES
TRACER TECHNIQUES
TRANSFERASES