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Effect of a single amino acid substitution G98D in a ribosome-associated essential GTPase, CgtA, on the growth and morphology of Vibrio cholerae

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Abstract

CgtA, a highly conserved 50S ribosome-associated essential GTPase, acts as a repressor of the stringent stress response under nutrient-rich growth conditions to suppress basal levels of the alarmone ppGpp in V. cholerae. To further explore the in vivo functionality of CgtA, we introduced an amino acid substitution, i.e., Gly98Asp, in a conserved glycine residue in the N-terminal domain. The constructed V. cholerae mutant was designated CgtA(G98D). Comparison of cell sizes of the CgtA(G98D)mutant with its isogenic wild-type (Wt) strain N16961 under different phases of growth by Transmission Electron Microscopy (TEM) and statistical analysis suggests that CgtA may control the cell size of V. cholerae. The cell length is significantly reduced, corresponding to the delayed growth in the mid-logarithmic phase. The differences in the cell length of CgtA(G98D) and Wt are indistinguishable in the late logarithmic phase. During the stationary phase, marked by higher OD600, a sub-population of CgtA(G98D) cells outnumbered the Wt cells lengthwise. CgtA(G98D) cells appeared slenderer than Wt cells with significantly reduced cell width. However, the centerline curvature is preserved in CgtA(G98D) cells. We propose that in addition to its multitude of intracellular roles, CgtA may influence the cell size of V. cholerae.

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Data availability

All requests for data, strains, and plasmids constructed in this study should be directed to the corresponding author.

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Acknowledgements

The authors acknowledge Dr. Rupak K. Bhadra from the Indian Institute of Chemical Biology, Kolkata, for providing the Wt Vibrio cholerae N16961 strain, Dr. Ananya Chatterjee from the ICMR-NICED, Kolkata, for helpful suggestions, Indian Institute of Science Education & Research (IISER), Kolkata for providing the instrumentation facility. A Ph.D. fellowship from IISER-Kolkata supports SD. The authors acknowledge DST-SERB of India (File Number: EMR/2015/002473) for providing research funds to PPD.

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Authors and Affiliations

  1. Department of Biological Sciences, Indian Institute of Science Education and Research (IISER), Mohanpur, Nadia, Kolkata, 741246, West Bengal, India

    Sagarika Das & Partha Pratim Datta

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  1. Sagarika Das
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  2. Partha Pratim Datta
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Concept, planning, experimental design, data analysis, and manuscript writing by PPD. Experimental design, execution, data analysis, and manuscript writing by SD.

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Correspondence to Partha Pratim Datta.

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Communicated by Erko Stackebrandt.

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Das, S., Datta, P.P. Effect of a single amino acid substitution G98D in a ribosome-associated essential GTPase, CgtA, on the growth and morphology of Vibrio cholerae. Arch Microbiol 204, 617 (2022). https://doi.org/10.1007/s00203-022-03233-w

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