Production of Mitotic Regulators Using an Autoselection System for Protein Expression in Budding Yeast

Author(s):  
Marco Geymonat ◽  
Adonis Spanos ◽  
Steven Sedgwick
2017 ◽  
Vol 156 ◽  
pp. 5-19 ◽  
Author(s):  
Emmanuelle Becker ◽  
Emmanuelle Com ◽  
Régis Lavigne ◽  
Marie-Hélène Guilleux ◽  
Bertrand Evrard ◽  
...  

2018 ◽  
Vol 29 (22) ◽  
pp. 2644-2655 ◽  
Author(s):  
Christina M. Kelliher ◽  
Matthew W. Foster ◽  
Francis C. Motta ◽  
Anastasia Deckard ◽  
Erik J. Soderblom ◽  
...  

In the budding yeast Saccharomyces cerevisiae, transcription factors (TFs) regulate the periodic expression of many genes during the cell cycle, including gene products required for progression through cell-cycle events. Experimental evidence coupled with quantitative models suggests that a network of interconnected TFs is capable of regulating periodic genes over the cell cycle. Importantly, these dynamical models were built on transcriptomics data and assumed that TF protein levels and activity are directly correlated with mRNA abundance. To ask whether TF transcripts match protein expression levels as cells progress through the cell cycle, we applied a multiplexed targeted mass spectrometry approach (parallel reaction monitoring) to synchronized populations of cells. We found that protein expression of many TFs and cell-cycle regulators closely followed their respective mRNA transcript dynamics in cycling wild-type cells. Discordant mRNA/protein expression dynamics was also observed for a subset of cell-cycle TFs and for proteins targeted for degradation by E3 ubiquitin ligase complexes such as SCF (Skp1/Cul1/F-box) and APC/C (anaphase-promoting complex/cyclosome). We further profiled mutant cells lacking B-type cyclin/CDK activity ( clb1-6) where oscillations in ubiquitin ligase activity, cyclin/CDKs, and cell-cycle progression are halted. We found that a number of proteins were no longer periodically degraded in clb1-6 mutants compared with wild type, highlighting the importance of posttranscriptional regulation. Finally, the TF complexes responsible for activating G1/S transcription (SBF and MBF) were more constitutively expressed at the protein level than at periodic mRNA expression levels in both wild-type and mutant cells. This comprehensive investigation of cell-cycle regulators reveals that multiple layers of regulation (transcription, protein stability, and proteasome targeting) affect protein expression dynamics during the cell cycle.


Yeast ◽  
2007 ◽  
Vol 24 (3) ◽  
pp. 145-154 ◽  
Author(s):  
Min-Woo Lee ◽  
Beom-Jun Kim ◽  
Hyun-Kyung Choi ◽  
Min-Jung Ryu ◽  
Sang-Bae Kim ◽  
...  

2010 ◽  
Vol 34 (8) ◽  
pp. S12-S12
Author(s):  
Hong‑Ge Li ◽  
Chen Min Xu ◽  
Kun Li ◽  
Ya Ni ◽  
Wen‑Ying Chen ◽  
...  

2000 ◽  
Vol 111 (4) ◽  
pp. 1118-1121 ◽  
Author(s):  
A. Bellahcene ◽  
I. Van Riet ◽  
C. de Greef ◽  
N. Antoine ◽  
M. F. Young ◽  
...  

2007 ◽  
Vol 177 (4S) ◽  
pp. 222-222
Author(s):  
Mireia Musquera ◽  
Maria J. Ribal ◽  
Yolanda Arce ◽  
Humberto Villavicencio ◽  
Fernando Algaba ◽  
...  

2006 ◽  
Vol 175 (4S) ◽  
pp. 323-323
Author(s):  
Gerald B. Brock ◽  
Ling De Young ◽  
Bing Siang Gan ◽  
Anthony J. Bella ◽  
Kok Bin Lim

2006 ◽  
Vol 175 (4S) ◽  
pp. 132-132
Author(s):  
Richard E. Zigeuner ◽  
Sebastian Leibl ◽  
Georg Hutterer ◽  
Thomas Chromecki ◽  
Peter Rehak ◽  
...  

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