Increase in the levels of activity of polyadenylic acid-metabolizing enzymes following phytohaemagglutinin stimulation of human lymphocytes

Increased levels of soluble activity of all three enzymes involved in polyadenylic acid metabolism were measured in PHA-stimulated versus normal lymphocytes. Poly(A)-polymerase and poly(A)-exonuclease values increased significantly (from 25.7 ± 4.2 (S.E.M.) to 53.5 ± 10.6 (S.E.M.), and from 334.6 ± 33.2 (S.E.M.) to 653.2 ± 53.4 (S.E. M.) respectively), while a moderate increase was observed in poly(A)-endonuclease (from 299.2 ± 33.8 (S.E.M.) to 403.0 ± 77.1 (S.E.M.). The above differences persisted after two fractionations of the crude cell extracts by ion exchange chromatography and molecular sieving, and could not be attributed to the competitive action of all three enzymes in the untreated extracts. Fractionation of the extracts of resting and stimulated cells on Sephadex G-75 revealed two molecular forms of poly(A)-polymerase activity.Abbreviations poly(A) or An Polyadenylic acid - oligo(A) or A₁₀ oligoadenylic acid - poly(C) polycytidylic acid - poly(U) polycytidylic acid - poly(G) polyguanylic acid - poly(dA) polydeoxyadenylic acid - EDTA ethylenediamine tetraacetate - PHA phytohaemagglutinin - PBS phosphate buffered saline - NP-40 nonidet-40 - KPi buffer potassium phosphate buffer