dictyNews
Electronic Edition
Volume 41, number 17
Sugust 14, 2015

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=========
Abstracts
=========


Proximity-dependent biotin identification (BioID) in 
Dictyostelium amoebae

Petros Batsios, Irene Meyer and Ralph Gräf

Institute for Biochemistry and Biology, Dept. of Cell Biology, 
University of Potsdam, D-14476 Potsdam-Golm, 
Karl-Liebknecht-Str. 24-25, 


Methods in Enzymology, in press

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 E. 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.


Submitted by Ralph Graef [[log in to unmask]] 
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[End dictyNews, volume 41, number 17]