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

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ADP-ribosylation and NAD+ Utilizing Enzymes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1813))

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.

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Acknowledgments

We thank members of the Lakin and Pears laboratories for constructive comments during the preparation of this manuscript. N.L.’s laboratory is supported by Cancer Research UK (www.cancerresearch.org.uk; grant C1521/A12353), Medical Research Council (www.mrc.ac.uk; MR/L000164/1), and NC3Rs (www.nc3rs.org.uk; NC/K00137X/1). AR was supported by a Clarendon Award (University of Oxford). C.P.’s laboratory is supported by NC3Rs (www.nc3rs.org.uk; NC/M000834/1).

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  1. Department of Biochemistry, University of Oxford, Oxford, UK

    Anna-Lena Kolb, Duen-Wei Hsu, Ana B. A. Wallis, Seiji Ura, Alina Rakhimova, Catherine J. Pears & Nicholas D. Lakin

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  1. Anna-Lena Kolb
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  2. Duen-Wei Hsu
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Correspondence to Nicholas D. Lakin .

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  1. Department of Biology, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA

    Paul Chang

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Kolb, AL. et al. (2018). Dictyostelium as a Model to Assess Site-Specific ADP-Ribosylation Events. In: Chang, P. (eds) ADP-ribosylation and NAD+ Utilizing Enzymes. Methods in Molecular Biology, vol 1813. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8588-3_9

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  • DOI: https://doi.org/10.1007/978-1-4939-8588-3_9

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