dictyNews Volume 41 Number 17

Published in

· 10 months ago

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 [rgraef@uni-potsdam.de]  
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[End dictyNews, volume 41, number 17]