@article {43051, title = {Tandem fluorescent protein timers for in vivo analysis of protein dynamics.}, journal = {Nature Biotechnology}, volume = {30}, year = {2012}, month = {2012 Jun 24}, pages = {708-14}, abstract = {

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{\textendash}mediated protein degradation.

}, keywords = {Green Fluorescent Proteins, High-Throughput Screening Assays, Kinetics, Nuclear Pore, Protein Stability, Proteins, Proteolysis, Saccharomyces cerevisiae, Subcellular Fractions}, issn = {1546-1696}, doi = {10.1038/nbt.2281}, author = {Khmelinskii, Anton and Keller, Philipp J and Bartosik, Anna and Meurer, Matthias and Barry, Joseph D and Mardin, Balca R and Kaufmann, Andreas and Trautmann, Susanne and Wachsmuth, Malte and Pereira, Gislene and Huber, Wolfgang and Schiebel, Elmar and Knop, Michael} } @article {1682, title = {Segregation of yeast nuclear pores.}, journal = {Nature}, volume = {466}, year = {2010}, month = {2010 Jul 22}, pages = {E1}, abstract = {

During mitosis in Saccharomyces cerevisiae, senescence factors such as extrachromosomal ribosomal DNA circles (ERCs) are retained in the mother cell and excluded from the bud/daughter cell. Shcheprova et al. proposed a model suggesting segregation of ERCs through their association with nuclear pore complexes (NPCs) and retention of preexisting NPCs in the mother cell during mitosis. However, this model is inconsistent with previous data and we demonstrate here that NPCs do efficiently migrate from the mother into the bud. Therefore, binding to NPCs does not seem to explain the retention of ERCs in the mother cell.

}, keywords = {Chromosome Segregation, DNA, Fungal, Mitosis, Nuclear Pore, Nuclear Pore Complex Proteins, Ribosomal, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins}, issn = {1476-4687}, doi = {10.1038/nature09255}, author = {Khmelinskii, Anton and Keller, Philipp J and Lorenz, Holger and Schiebel, Elmar and Knop, Michael} }