Influence of phosphate on activity and stability of reverse transcriptase from avian myeloblastosis virus.
Activity of RNA-dependent DNA polymerase (RDDP) from avian myeloblastosis virus (AMV), either in purified form or in virus lysates, was increased by phosphorylation. Stability of RDDP in lysates buffered with phosphate was much greater (no loss of activity in 48 hours at 4 degrees) than that in lysates buffered with Tris-Cl (76% loss). Activity lost in the Tris-buffered extracts was completely restored by phosphorylation. The findings suggested that AMV RDDP activity is influenced by the degree of phosphorylation of the enzyme or enzyme-associated proteins and that this chemical modification is mediated by protein phosphokinase and phosphoprotein phosphatase present in crude extracts of purified AMV. Application of these results provided the basis of procedures whereby RDDP can be recovered in significantly higher yield and purity than formerly.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000469:Alkaline Phosphatase / Q000494:pharmacology; D001354:Avian Leukosis Virus / Q000201:enzymology*; D009189:Avian Myeloblastosis Virus / Q000201:enzymology*; D002021:Buffers; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D010750:Phosphoproteins / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*
D002021:Buffers; D009943:Organophosphorus Compounds; D010750:Phosphoproteins; D000255:Adenosine Triphosphate; D012194:RNA-Directed DNA Polymerase; D000469:Alkaline Phosphatase