Immunolocalization of Poly ADP-Ribose on Drosophila Polytene Chromosomes

Poly ADP-ribosylation (PARylation) is a posttranslational protein modification catalyzed by poly �ADP-ribose polymerases (PARPs). Poly ADP-ribose metabolism is involved in a wide range of biological processes, such as maintenance of genome stability, transcriptional regulation, energy metabolism, and programed cell death. Recently, chromatin components, including histones, have been shown to be targets of PARylation. Unlike mammals, which have several PARP-encoded genes, the model organism Drosophila melanogaster has only one PARP gene, highly related to mammalian PARP1. These features make flies a great model system to study PARP biology. Commercially available antibodies recognizing this covalent modification have made possible the development of immunofluorescence approaches to study PARylation of chromatin components. Here, we present a protocol to immunostain polytene chromosomes of the model system D. melanogaster .