Phosphorylation and immunology of poly(A) polymerase

Abstract

Phosphorylation of poly(A) polymerase by protein kinase NI (a cyclic nucleotide-independent nuclear kinase closely associated with poly(A) polymerase at early stages of purification) resulted in as much as 7-fold activation of poly(A) polymerase. Phosphorylation causes an increase in the rate rather than the extent of polyadenylation. Antibodies raised in rabbits against purified poly(A) polymerase from Morris hepatoma 3924A reacted specifically with poly(A) polymerase following “Western” transfer of the enzyme onto diazobenzyloxyl methyl paper. Using iodinated enzyme, a competition radioimmunoassay for poly(A) polymerase was developed. Using the radioimmunoassay, it was shown that Morris hepatoma 3924A contains 100 μg of poly(A) polymerase/mg DNA or 10 7 molecules of the enzyme/cell nucleus. Nuclear poly(A) polymerase from fetal liver, but not from normal liver, was able to compete well with hepatoma enzyme in the radioimmunoassay. These data suggest that the tumor poly(A) polymerase is probably an oncofetal antigen, resulting from derepression of a gene not normally expressed in adult liver.