DOI: 10.1128/jb.177.6.1497-1504.1995. Gratuitous overexpression of genes in Escherichia coli leads to growth inhibition and ribosome destruction

Gratuitous overexpression of genes in Escherichia coli leads to growth inhibition and ribosome destruction

10.1128/jb.177.6.1497-1504.1995

Crossref journal-article

American Society for Microbiology

Journal of Bacteriology (235)

Abstract

We attempted to test the idea that the relative abundance of each individual tRNA isoacceptor in Escherichia coli can be altered by varying its cognate codon concentration. In order to change the overall codon composition of the messenger pool, we have expressed in E. coli lacZ with the aid of T7 RNA polymerase so that their respective gene products individually accounted for 30% of the total bacterial protein. Unexpectedly, the maximum expression of either test gene has no specific effect on the relative rates of synthesis of the tRNA species that we studied. Instead, we find that there is a cumulative breakdown of rRNAs, which results in a loss of ribosomes and protein synthetic capacity. After either of the test genes is maximally induced, there is a growing fraction of protein synthesis invested in beta-galactosidase or delta tufB that is matched by a comparable decrease of the fraction of normal protein synthesis. We have also observed enhanced accumulation of two heat shock proteins during overexpression. Finally, after several hours of overexpression of either test protein, the bacteria are no longer viable. These results are relevant to the practical problems of obtaining high expression levels for cloned proteins.

Bibliography

Dong, H., Nilsson, L., & Kurland, C. G. (1995). Gratuitous overexpression of genes in Escherichia coli leads to growth inhibition and ribosome destruction. Journal of Bacteriology, 177(6), 1497â1504.

Authors 3

H Dong (first)
L Nilsson (additional)
C G Kurland (additional)

References 27 Referenced 297

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Emilsson , V. , and C. G. Kurland . 1990 . Growth rate dependence of transfer RNA abundance in Escherichia coli . EMBO J. 9 : 4359 - 4366 . (10.1002/j.1460-2075.1990.tb07885.x) / EMBO J. / Growth rate dependence of transfer RNA abundance in Escherichia coli by Emilsson V. (1990)
Emilsson , V. , and C. G. Kurland . 1990 . Growth rate dependence of global amino acid composition . Biochim. Biophys. Acta 1050 : 248 - 251 . (10.1016/0167-4781(90)90175-2) / Biochim. Biophys. Acta / Growth rate dependence of global amino acid composition by Emilsson V. (1990)
Emilsson , V. , A. K. Näslund , and C. G. Kurland . 1993 . Growth rate dependent accumulation of twelve tRNA species in Escherichia coli . J. Mol. Biol. 230 : 483 - 491 . (10.1006/jmbi.1993.1165) / J. Mol. Biol. / Growth rate dependent accumulation of twelve tRNA species in Escherichia coli by Emilsson V. (1993)
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Gausing , K. 1977 . Regulation of ribosome production in Escherichia coli: synthesis and stability of ribosomal RNA and of ribosomal protein messenger RNA at different growth rates . J. Mol. Biol. 115 : 335 - 354 . (10.1016/0022-2836(77)90158-9) / J. Mol. Biol. / Regulation of ribosome production in Escherichia coli: synthesis and stability of ribosomal RNA and of ribosomal protein messenger RNA at different growth rates by Gausing K. (1977)
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Ikemura , T. 1981 . Correlation between the abundance of Escherichia coli transfer RNAs and the occurrence of the respective codons in its protein genes . J. Mol. Biol. 146 : 1 - 21 . (10.1016/0022-2836(81)90363-6) / J. Mol. Biol. / Correlation between the abundance of Escherichia coli transfer RNAs and the occurrence of the respective codons in its protein genes by Ikemura T. (1981)
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Pedersen S. 1976. Stability of nascent ribosomal RNA in Escherichia coli p. 345-355. In N. O. Kjelgaard and O. Maaløe (ed.) Control of ribosome synthesis. Alfred Benzon Symposium. IX. Munksgaard Copenhagen.
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Dates

Type	When
Created	8 years, 9 months ago (Nov. 9, 2016, 11:23 a.m.)
Deposited	4 years ago (July 29, 2021, 1:55 p.m.)
Indexed	1 month ago (July 26, 2025, 5:07 a.m.)
Issued	30 years, 5 months ago (March 1, 1995)
Published	30 years, 5 months ago (March 1, 1995)
Published Print	30 years, 5 months ago (March 1, 1995)

Funders 0

None

BibTeX

@article{Dong_1995, title={Gratuitous overexpression of genes in Escherichia coli leads to growth inhibition and ribosome destruction}, volume={177}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.177.6.1497-1504.1995}, DOI={10.1128/jb.177.6.1497-1504.1995}, number={6}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Dong, H and Nilsson, L and Kurland, C G}, year={1995}, month=mar, pages={1497–1504} }

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