Purification and Properties of Poly(A) Polymerase from Vigna unguiculata

Poly(A) polymerase was purified from germinating Vigna unguiculataseeds by successive column chromatography on phosphocellulose,Toyopearl HW-55S, heparin-Sepharose and TSKgel phenyl-5PW, whichyielded two activity fractions. The first fraction was purifiedas a single polypeptide with a mol wt of 63,000 as estimatedby SDS-PAGE. The enzyme activity was highly specific for ATPand required Mn²⁺ ion; an ATP-Mn complex may be the actual substrate.The polymerization reaction required a primer, with varioustypes of RNAs, poly(A) as well as dinucleoside phosphates having3'—OH, serving as efficient primers. The two forms ofthe enzyme had very similar properties with respect to divalentcation requirement and dependency on ion strength, but theyshowed some difference in primer preference. (Received March 4, 1988; Accepted May 2, 1988)