Journal
NATURE BIOTECHNOLOGY
Volume 30, Issue 7, Pages 708-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nbt.2281
Keywords
-
Categories
Funding
- Cell Networks cluster
- German Research Foundation [SFB638]
- Howard Hughes Medical Institute
- European Molecular Biology Organization [EMBO ALTF 1124-2010]
- EU-FP7 Network of Excellence in Systems Microscopy
- Novartis Stiftung
Ask authors/readers for more resources
The functional state of a cell is largely determined by the spatiotemporal organization of its proteome. Technologies exist for measuring particular aspects of protein turnover and localization, but comprehensive analysis of protein dynamics across different scales is possible only by combining several methods. Here we describe tandem fluorescent protein timers (tFTs), fusions of two single-color fluorescent proteins that mature with different kinetics, which we use to analyze protein turnover and mobility in living cells. We fuse tFTs to proteins in yeast to study the longevity, segregation and inheritance of cellular components and the mobility of proteins between subcellular compartments; to measure protein degradation kinetics without the need for time-course measurements; and to conduct high-throughput screens for regulators of protein turnover. Our experiments reveal the stable nature and asymmetric inheritance of nuclear pore complexes and identify regulators of N-end rule-mediated protein degradation.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available