A Chemostat Array Enables the Spatio-Temporal Analysis of the Yeast Proteome
N. Dénervaud, J. Becker, R. Delgado-Gonzalo, P. Damay, A.S. Rajkumar, M. Unser, D. Shore, F. Naef, S.J. Maerki
Proceedings of the National Academy of Sciences of the United States of America, vol. 110, no. 39, pp. 15842–15847, September 24, 2013.
Observing cellular responses to perturbations is central to generating and testing hypotheses in biology. We developed a massively parallel microchemostat array capable of growing and observing 1,152 yeast-GFP strains on the single-cell level with 20 min time resolution. We measured protein abundance and localization changes in 4,085 GFP-tagged strains in response to methyl methanesulfonate and analyzed 576 GFP strains in five additional conditions for a total of more than 10,000 unique experiments, providing a systematic view of the yeast proteome in flux. We observed that processing bodies formed rapidly and synchronously in response to UV irradiation, and in conjunction with 506 deletion-GFP strains, identified four gene disruptions leading to abnormal ribonucleotide-diphosphate reductase (Rnr4) localization. Our microchemostat platform enables the large-scale interrogation of proteomes in flux and permits the concurrent observation of protein abundance, localization, cell size, and growth parameters on the single-cell level for thousands of microbial cultures in one experiment.
@ARTICLE(http://bigwww.epfl.ch/publications/denervaud1301.html,
AUTHOR="D{\'{e}}nervaud, N. and Becker, J. and Delgado-Gonzalo, R. and
Damay, P. and Rajkumar, A.S. and Unser, M. and Shore, D. and Naef,
F. and Maerki, S.J.",
TITLE="A Chemostat Array Enables the Spatio-Temporal Analysis of the
Yeast Proteome",
JOURNAL="Proceedings of the National Academy of Sciences of the United
States of America",
YEAR="2013",
volume="110",
number="39",
pages="15842--15847",
month="September 24,",
note="")