Format

Send to

Choose Destination
Methods Mol Biol. 2018;1813:125-148. doi: 10.1007/978-1-4939-8588-3_9.

Dictyostelium as a Model to Assess Site-Specific ADP-Ribosylation Events.

Author information

1
Department of Biochemistry, University of Oxford, Oxford, UK.
2
Department of Biochemistry, University of Oxford, Oxford, UK. nicholas.lakin@bioch.ox.ac.uk.

Abstract

The amoeba Dictyostelium discoideum is a single-cell organism that can undergo a simple developmental program, making it an excellent model to study the molecular mechanisms of cell motility, signal transduction, and cell-type differentiation. A variety of human genes that are absent or show limited conservation in other invertebrate models have been identified in this organism. This includes ADP-ribosyltransferases, also known as poly-ADP-ribose polymerases (PARPs), a family of proteins that catalyze the addition of single or poly-ADP-ribose moieties onto target proteins. The genetic tractability of Dictyostelium and its relatively simple genome structure makes it possible to disrupt PARP gene combinations, in addition to specific ADP-ribosylation sites at endogenous loci. Together, this makes Dictyostelium an attractive model to assess how ADP-ribosylation regulates a variety of cellular processes including DNA repair, transcription, and cell-type specification. Here we describe a range of techniques to study ADP-ribosylation in Dictyostelium, including analysis of ADP-ribosylation events in vitro and in vivo, in addition to approaches to assess the functional roles of this modification in vivo.

KEYWORDS:

ADP-ribosylation; ADP-ribosyltransferase; DNA repair; Dictyostelium; PARP

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center