RT Journal Article
SR Electronic
T1 Systematic assessment of GFP tag position on protein localization and growth fitness in yeast
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 360636
DO 10.1101/360636
A1 Dan Davidi
A1 Uri Weill
A1 Gat Krieger
A1 Zohar Avihou
A1 Ron Milo
A1 Maya Schuldiner
YR 2018
UL http://biorxiv.org/content/early/2018/07/03/360636.abstract
AB While protein tags are ubiquitously utilized in molecular biology, they harbor the potential to interfere with functional traits of their fusion counterparts. Systematic evaluation of the effect of protein tags on localization and function would promote accurate use of tags in experimental setups. Here we examine the effect of Green Fluorescent Protein (GFP) tagging at either the N or C terminus of budding yeast proteins on localization and functionality. We use a competition-based approach to decipher the relative fitness of two strains tagged on the same protein but on opposite termini and from that infer the correct, physiological localization for each protein and the optimal position for tagging. Our study provides a first of a kind systematic assessment of the effect of tags on the functionality of proteins and provides step towards broad investigation of protein fusion libraries.HighlightsProtein tags are widely used in molecular biology although they may interfere with protein function.The subcellular localization of hundreds of proteins in yeast is different when tagged at the N or the C terminus.A competition based assay enables systematic deciphering of correct tagging terminus for essential proteins.The presented approach can be used to derive physiologically relevant tagged libraries.