Resolution of DNA polymerase-. cap alpha. and primase activities from embryonic chicken brain and stimulation of chain initiation by a protein factor

Conference · Fri May 01 04:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5904489

Ray, S; Torres-Rosado, A; Basu, S

Reconstitution of an in vitro system capable of DNA chain initiation in an eukaryotic system has been attempted in several laboratories. They have previously reported the isolation of three distinct forms of DNA polymerase-..cap alpha.. from IMR-32 cells and 9-day-old embryonic chicken brains (9ECB). DNA primase activity is eluted from a DE-23 column with one of these Pol-..cap alpha..'s when a linear gradient is used. Irrespective of embryonic age, primase and a pol-..cap alpha.. activity coelutes at 250 mM K-PO/sub 4/ buffer. The complex containing Pol-..cap alpha../primase activities has been analyzed further on a continuous velocity gradient of either sucrose or glycerol. A polymerase-..cap alpha..-free primase activity sediments at 4.5S whereas the other 50% is recovered with the pol-..cap alpha.. activity which sediments at 11S. Compared to polydC and poly(dC dT), of the 9ECB primase catalyzed /sup 3/H-dXMP incorporation into polydT and M13 is very low. The template poly(dC, dT) is active only in the presence of ATP. They have purified a protein factor (NPF-1) from rat liver which stimulates primase-associated pol-..cap alpha.. activities from IMR-32 cells. The NPF-1 also stimulates (2 to 3-fold) primase-associated pol-..cap alpha.. activity (11S) from 9ECB.

• Research Organization: Univ. of Notre Dame, IN
• OSTI ID: 5904489
• Report Number(s): CONF-870644-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 46:6
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BIOCHEMICAL REACTION KINETICS
BIRDS
BODY
BRAIN
CENTRAL NERVOUS SYSTEM
CHICKENS
DNA POLYMERASES
EMBRYOS
ENZYME ACTIVITY
ENZYMES
FOWL
FRACTIONATION
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
NERVOUS SYSTEM
NUCLEOTIDYLTRANSFERASES
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PROTEINS
REACTION KINETICS
RNA POLYMERASES
SEPARATION PROCESSES
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES