TY  - JOUR
AU  - Lazarewicz, Natalia
AU  - Le Dez, Gaëlle
AU  - Cerjani, Romina
AU  - Runeshaw, Lunelys
AU  - Meurer, Matthias
AU  - Knop, Michael
AU  - Wysocki, Robert
AU  - Rabut, Gwenaël
TI  - Accurate and sensitive interactome profiling using a quantitative protein-fragment complementation assay.
JO  - Cell reports / Methods
VL  - 4
IS  - 10
SN  - 2667-2375
CY  - Cambridge, MA
PB  - Cell Press
M1  - DKFZ-2024-02167
SP  - 100880
PY  - 2024
N1  - DKFZ-ZMBH Alliance
AB  - An accurate description of protein-protein interaction (PPI) networks is key to understanding the molecular mechanisms underlying cellular systems. Here, we constructed genome-wide libraries of yeast strains to systematically probe protein-protein interactions using NanoLuc Binary Technology (NanoBiT), a quantitative protein-fragment complementation assay (PCA) based on the NanoLuc luciferase. By investigating an array of well-documented PPIs as well as the interactome of four proteins with varying levels of characterization-including the well-studied nonsense-mediated mRNA decay (NMD) regulator Upf1 and the SCF complex subunits Cdc53 and Met30-we demonstrate that ratiometric NanoBiT measurements enable highly precise and sensitive mapping of PPIs. This work provides a foundation for employing NanoBiT in the assembly of more comprehensive and accurate protein interaction maps as well as in their functional investigation.
KW  - Protein Interaction Mapping: methods
KW  - Saccharomyces cerevisiae: genetics
KW  - Saccharomyces cerevisiae: metabolism
KW  - Protein Interaction Maps
KW  - Saccharomyces cerevisiae Proteins: metabolism
KW  - Saccharomyces cerevisiae Proteins: genetics
KW  - Luciferases: genetics
KW  - Luciferases: metabolism
KW  - CP: Systems biology (Other)
KW  - Cdc53 (Other)
KW  - Irc20 (Other)
KW  - Met30 (Other)
KW  - Nam7 (Other)
KW  - NanoBiT (Other)
KW  - Saccharomyces cerevisiae (Other)
KW  - Upf1 (Other)
KW  - budding yeast (Other)
KW  - interactome (Other)
KW  - protein-protein interaction (Other)
KW  - Saccharomyces cerevisiae Proteins (NLM Chemicals)
KW  - Luciferases (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:39437715
DO  - DOI:10.1016/j.crmeth.2024.100880
UR  - https://inrepo02.dkfz.de/record/294334
ER  -