TY - JOUR
T1 - Recombination-induced tag exchange to track old and new proteins
AU - Verzijlbergen, Kitty F.
AU - Menendez-Benito, Victoria
AU - van Welsem, Tibor
AU - van Deventer, Sjoerd J.
AU - Lindstrom, Derek L.
AU - Ovaa, Huib
AU - Neefjes, Jacques
AU - Gottschling, Daniel E.
AU - van Leeuwen, Fred
PY - 2010
Y1 - 2010
N2 - The dynamic behavior of proteins is critical for cellular homeostasis. However, analyzing dynamics of proteins and protein complexes in vivo has been difficult. Here we describe recombination-induced tag exchange (RITE), a genetic method that induces a permanent epitope-tag switch in the coding sequence after a hormone-induced activation of Cre recombinase. The time-controlled tag switch provides a unique ability to detect and separate old and new proteins in time and space, which opens up opportunities to investigate the dynamic behavior of proteins. We validated the technology by determining exchange of endogenous histones in chromatin by biochemical methods and by visualizing and quantifying replacement of old by new proteasomes in single cells by microscopy. RITE is widely applicable and allows probing spatiotemporal changes in protein properties by multiple methods.
AB - The dynamic behavior of proteins is critical for cellular homeostasis. However, analyzing dynamics of proteins and protein complexes in vivo has been difficult. Here we describe recombination-induced tag exchange (RITE), a genetic method that induces a permanent epitope-tag switch in the coding sequence after a hormone-induced activation of Cre recombinase. The time-controlled tag switch provides a unique ability to detect and separate old and new proteins in time and space, which opens up opportunities to investigate the dynamic behavior of proteins. We validated the technology by determining exchange of endogenous histones in chromatin by biochemical methods and by visualizing and quantifying replacement of old by new proteasomes in single cells by microscopy. RITE is widely applicable and allows probing spatiotemporal changes in protein properties by multiple methods.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=76249091039&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/20018668
U2 - https://doi.org/10.1073/pnas.0911164107
DO - https://doi.org/10.1073/pnas.0911164107
M3 - Article
C2 - 20018668
SN - 0027-8424
VL - 107
SP - 64
EP - 68
JO - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
JF - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
IS - 1
ER -