Association of diadenosine 5‘,5“‘-P1,P4-tetraphosphate binding protein with HeLa cell DNA polymerase alpha.

Abstract

An electrophoretically homogeneous high molecular weight form (640,000) of HeLa cell DNA polymerase alpha was shown to catalyze DNA synthesis with a variety of di- and oligoriboadenylates and oligodeoxyriboadenylates as primers with poly(dT) as template. Diadenosine 5',5"'-P1,p4-tetraphosphate (Ap4A) can be utilized as a primer with poly(dT) as template and was found to be covalently attached to the 5'-end of the poly(dA) product. An Ap4A binding protein is tightly associated with the high molecular weight form of DNA polymerase alpha. This protein which exhibits high affinity, noncovalent binding of Ap4A is resolved from the multiprotein DNA polymerase alpha complex, along with other accessory proteins, by hydrophobic affinity chromatography on phenyl-Sepharose columns.