Proximity-Dependent Biotin Identification (BioID) in Dictyostelium Amoebae

Methods in Enzymology
Petros BatsiosRalph Gräf

Abstract

The identification of a bona fide lamin-like protein in Dictyostelium made this lower eukaryote an attractive model organism to study evolutionarily conserved nuclear envelope (NE) proteins important for nuclear organization and human laminopathies. Proximity-dependent biotin identification (BioID), reported by Roux and colleagues, is a powerful discovery tool for lamin-associated proteins. In this method, living cells express a bait protein (e.g., lamin) fused to an R118G-mutated version of BirA, an Escherichia coli biotinylase. In the presence of biotin, BirA-R118G biotinylates target proteins in close proximity in vivo, which are purified using streptavidin and identified by immunoblotting or mass spectrometry. We adapted the BioID method for use in Dictyostelium amoebae. The protocols described here successfully revealed Dictyostelium lamin-like protein NE81 proximity to Sun1, a conserved inner nuclear membrane protein.

Citations

Jan 21, 2017·European Journal of Cell Biology·Irene MeyerRalph Gräf
Mar 9, 2018·Current Protocols in Protein Science·Kyle J RouxDanielle G May
Feb 20, 2019·Cells·Marianne GrafeRalph Gräf
Feb 19, 2019·F1000Research·Laura Trinkle-Mulcahy
Jan 13, 2021·Nature Methods·Wei QinAlice Y Ting
Aug 1, 2021·Molecular & Cellular Proteomics : MCP·Shimon M RosenthalAnne-Claude Gingras

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