A chemostat array enables the spatio-temporal analysis of the yeast proteome

被引：95

作者：

Denervaud, Nicolas ^{[1
]}

Becker, Johannes ^{[1
]}

Delgado-Gonzalo, Ricard ^{[2
]}

Damay, Pascal ^{[3
,4
]}

Rajkumar, Arun S. ^{[1
]}

Unser, Michael ^{[2
]}

Shore, David ^{[3
,4
]}

Naef, Felix ^{[1
]}

Maerkl, Sebastian J. ^{[1
]}

机构：

[1] Ecole Polytech Fed Lausanne, Inst Bioengn, CH-1015 Lausanne, Switzerland

[2] Ecole Polytech Fed Lausanne, Sch Engn, CH-1015 Lausanne, Switzerland

[3] Univ Geneva, Dept Mol Biol, CH-1211 Geneva, Switzerland

[4] Univ Geneva, Natl Ctr Competence Res NCCR Program Frontiers Ge, CH-1211 Geneva, Switzerland

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2013年 / 110卷 / 39期

关键词：

microfluidics; cell arrays; high-content imaging; DNA damage response; yeast proteomics; DNA-DAMAGE CHECKPOINT; SACCHAROMYCES-CEREVISIAE; GLOBAL ANALYSIS; BUDDING YEAST; SINGLE CELLS; P-BODIES; LOCALIZATION; REVEALS; REPLICATION; EXPRESSION;

D O I：

10.1073/pnas.1308265110

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：15842 / 15847

页数：6

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