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. 2021 Jul 26:8:669996.
doi: 10.3389/fmolb.2021.669996. eCollection 2021.

Chaperonin Abundance Enhances Bacterial Fitness

C M Santosh Kumar  1 , Kritika Chugh  2 , Anirban Dutta  3 , Vishnuvardhan Mahamkali  4 , Tungadri Bose  3 , Sharmila S Mande  3 , Shekhar C Mande  5 , Peter A Lund  1
Affiliations

Chaperonin Abundance Enhances Bacterial Fitness

C M Santosh Kumar et al. Front Mol Biosci. .

Abstract

The ability of chaperonins to buffer mutations that affect protein folding pathways suggests that their abundance should be evolutionarily advantageous. Here, we investigate the effect of chaperonin overproduction on cellular fitness in Escherichia coli. We demonstrate that chaperonin abundance confers 1) an ability to tolerate higher temperatures, 2) improved cellular fitness, and 3) enhanced folding of metabolic enzymes, which is expected to lead to enhanced energy harvesting potential.

Keywords: GroEL; competitive index; evolution; metabolic flux; metabolism; proteomics.

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Conflict of interest statement

Authors AD, TB, and SSM are employed by the company TCS Research division in the Tata Consultancy Services Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Construction of GL-Lt and GL-Ht Strains. In E. coli LG6, the groE operon is under the control of the inducible P trc promoter. This strain was transformed with either a plasmid expressing groE under the control of lactose (GroE Plasmid), pTrc-GSL or its empty vector (Vector), pTrc99a. Upon culturing in the presence of lactose, GL-Ht produces elevated levels of GroEL and GroES, due to the induction of chromosomal and plasmid-borne groE operon, while GL-Lt will have lower production of GroES and GroEL due to the induction of only the chromosomal groE copy.
FIGURE 2
FIGURE 2
Temperature Tolerance upon Chaperonin Overproduction. Ten-fold serially diluted (10−1 through 10−6) exponentially growing cultures of GL-Ht (H), GL-Lt (L), MG16155 (W), and MG1655 with pTrc-GSL (W+) were spotted onto LB agar plates supplemented with lactose. These plates were incubated at the indicated temperatures.
FIGURE 3
FIGURE 3
Chaperonin Depletion leads to Lower CI (A) Strategy for Determining the CI for GL-Lb and GL-Hb strains. Stationary phase cultures of GL-Lb and GL-Hb strains were mixed at equal cell density, grown to stationary phase and sub-cultured in fresh media for 20 continuous passages. Cells recovered at each passage were serially diluted as indicated, spread on LB agar plates and the resulting colonies were scored for their phenotype (GL-Lb or GL-Hb), by colony PCR, using vector specific oligonucleotide primers (B) CI, as a degree of fitness, was calculated at every passage from a ratio of proportion of the cells with a particular phenotype and plotted as a function of number of passages. CI trend was similar among the three independent experiments.
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