Evaluation of the lower protein limit in the budding yeast Saccharomyces cerevisiae using TIPI-gTOW.

ABSTRACT:

Background

Identifying permissible limits of intracellular parameters such as protein expression provides important information for examining robustness. In this study, we used the TEV protease-mediated induction of protein instability (TIPI) in combination with the genetic Tug-of-War (gTOW) to develop a method to measure the lower limit of protein level. We first tested the feasibility of this method using ADE2 as a marker and then analyzed some cell cycle regulators to reveal genetic interactions.

Results

Using TIPI-gTOW, we successfully constructed a strain in which GFP-(TDegF)Ade2 was expressed at the lower limit, just sufficient to support cellular growth under the -Ade condition by accelerating degradation by TEV protease. We also succeeded in constructing a strain in

SUBMITTER: Sasabe M 

PROVIDER: S-EPMC3892067 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature