Nuclear transport of adenovirus DNA polymerase is facilitated by interaction with preterminal protein

Abstract

The mRNAs for the 80 kd adenovirus preterminal protein (pTP) and the 140 kd DNA polymerase (AdPol) contain several exons spliced to the main open reading frames (m-ORFs) located in the early transcription unit E2B. These proteins were transiently expressed in monkey kidney cells (CV1) utilizing the first ATG (pTP1 and AdPol1) or the ATG of a linker inserted at the beginning of the m-ORFs (pTP2 and AdPol2). Only pTP2 and AdPol2 were functionally active in an in vitro replication initiation assay. Both pTP1 and pTP2 were transported to the nucleus. The sequence RLPV(R)6VP, which is present in both pTPs, is identified as their nuclear localization signal. In contrast, AdPol1 was cytoplasmically localized, whereas AdPol2 was distributed in both compartments, suggesting that the nuclear localization signal for AdPol is within the first 139 amino acids. Interestingly, when AdPol1 and pTP1 or AdPol2 and pTP2 were coexpressed in the transfected cells, the nuclear distribution of AdPol1 or AdPol2 was significantly increased. We demonstrate that the nuclear transport of AdPol is facilitated, irrespective of the presence of its nuclear localization signal, by interaction with pTP.