ADP-ribosylation of nonhistone proteins during the HeLa cell cycle

ADP-ribosylation of nonhistone proteins during the HeLa cell cycle was investigated. Proteins were radiolabeled by incubating interphase nuclei and mitotic cells with the specific precursor, [32P]NAD. Autoradiograms of two-dimensional gels of total nuclear nonhistone proteins showed a large number of modified species (more than 140). A complex pattern was also found for interphase chromatin. Nuclear scaffolds showed a simpler pattern of four major groups of modified species, which appeared to be the lamins and poly(ADP-ribose) polymerase. The labeling pattern for nonhistones of metaphase chromosomes was fundamentally different than with interphase nuclei. Autoradiograms were dominated by the incorporation of label into poly(ADP-ribose) polymerase.