TY - JOUR
T1 - Estimating the protein burden limit of yeast cells by measuring the expression limits of glycolytic proteins
AU - Eguchi, Yuichi
AU - Makanae, Koji
AU - Hasunuma, Tomohisa
AU - Ishibashi, Yuko
AU - Kito, Keiji
AU - Moriya, Hisao
N1 - Funding Information:
We thank the members of the Moriya laboratories for advice and helpful discussions, Mr. Katsuhiro Yamamoto and Ms. Yoshimi Hori for experimental support, and Dr. Kei Takahashi for providing experimental materials. New energy and industrial technology development organization development of production techniques for highly functional biomaterials using smart cell, p16009 hisao moriya japan society for the promo- tion of science kakenhi 17h03618 hisao moriya japan society for the promo- tion of science kakenhi 15kk0258 hisao moriya the funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication
Publisher Copyright:
© Eguchi et al.
PY - 2018/8/10
Y1 - 2018/8/10
N2 - The ultimate overexpression of a protein could cause growth defects, which are known as the protein burden. However, the expression limit at which the protein-burden effect is triggered is still unclear. To estimate this limit, we systematically measured the overexpression limits of glycolytic proteins in Saccharomyces cerevisiae. The limits of some glycolytic proteins were up to 15% of the total cellular protein. These limits were independent of the proteins’ catalytic activities, a finding that was supported by an in silico analysis. Some proteins had low expression limits that were explained by their localization and metabolic perturbations. The codon usage should be highly optimized to trigger the protein-burden effect, even under strong transcriptional induction. The S–S-bond-connected aggregation mediated by the cysteine residues of a protein might affect its expression limit. Theoretically, only non-harmful proteins could be expressed up to the protein-burden limit. Therefore, we established a framework to distinguish proteins that are harmful and non-harmful upon overexpression.
AB - The ultimate overexpression of a protein could cause growth defects, which are known as the protein burden. However, the expression limit at which the protein-burden effect is triggered is still unclear. To estimate this limit, we systematically measured the overexpression limits of glycolytic proteins in Saccharomyces cerevisiae. The limits of some glycolytic proteins were up to 15% of the total cellular protein. These limits were independent of the proteins’ catalytic activities, a finding that was supported by an in silico analysis. Some proteins had low expression limits that were explained by their localization and metabolic perturbations. The codon usage should be highly optimized to trigger the protein-burden effect, even under strong transcriptional induction. The S–S-bond-connected aggregation mediated by the cysteine residues of a protein might affect its expression limit. Theoretically, only non-harmful proteins could be expressed up to the protein-burden limit. Therefore, we established a framework to distinguish proteins that are harmful and non-harmful upon overexpression.
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U2 - 10.7554/eLife.34595
DO - 10.7554/eLife.34595
M3 - Article
C2 - 30095406
AN - SCOPUS:85053884085
VL - 7
JO - eLife
JF - eLife
SN - 2050-084X
M1 - e34595
ER -