Adenovirus protein V is associated with the DNA-containing virus core and functions as a bridge between the capsid and the core. A yeast two-hybrid analysis performed with a human cDNA library using protein V as 'bait' selected a cellular protein, p32 --described previously as associated with the splicing factor ASF/SF2. By expression and purification of p32 and preparation of an antibody we confirmed the binding of p32 to V by a variety of methods including immune precipitation. We demonstrated that p32 was primarily located in the cytoplasm in association with mitochondria but could also be detected in the nucleus as distinct granules and tubules. By examining infected cells using confocal microscopy and immunofluorescence we were able to follow the intracellular locations of protein V and p32 and it is postulated that p32 is part of a system which imports proteins to the nucleus and that adenovirus hijacks this process to deliver its genome to the nucleus.