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Involvement of BglG in Lipopolysaccharides (LPS) Synthesis and Transport in Stationary Phase in E. coli

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Abstract

BglG, an RNA binding regulatory protein encoded by the β-glucoside (bgl) operon of E. coli is known to be involved in the regulation of several metabolic functions in stationary phase. A genome-wide comparative transcriptome analysis performed earlier between a ∆bglG strain and its isogenic WT counterpart revealed that genes involved in lipopolysaccharide (LPS) biosynthesis and transport were significantly down-regulated in the absence of BglG in stationary phase, suggesting a role for BglG in their regulation. We have investigated the involvement of BglG in LPS biosynthesis and transport. Consistent with the down-regulation of LPS synthesis and transport genes, the ∆bglG strain showed a loss of permeability barrier specifically in stationary phase, which could be rescued by introduction of wild type bglG on a plasmid. A search for a putative transcription factor involved in the regulation mediated by BglG led to the identification of GadE, which is one of the primary positive regulators of pH homeostasis and LPS core biosynthesis. Using RNA mobility shift and stability assays, we show that BglG binds specifically to gadE mRNA and enhances its stability. Consistent with this, loss of gadE leads to a partial defect in permeability. Based on our findings, we propose a model for the molecular mechanism involved in the regulation on LPS synthesis and transport by BglG.

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Abbreviations

LPS:

Lipopolysaccharide

PTS:

Phosphotransferase system

RAT:

Ribonucleic antiterminator

GAD:

Glutamate dependent

RNA EMSA:

Ribonucleic acid electrophoretic mobility shift assay

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Acknowledgements

This study was funded by the Department of Biotechnology (DBT) through a partnership programme with the Indian Institute of Science. The authors are also grateful to the Department of Science and Technology (DST) and the Universities Grants Commission (UGC) for infrastructural support. KV and SS are recipients of research fellowship from the Institute. We also thank the two anonymous referees for their constructive comments.

Funding

This study was funded by the Department of Biotechnology (DBT), Government of India, through a partnership program with the Indian Institute of Science. Infrastructural support was also provided by the Department of Science and Technology (DST) and the Universities Grants Commission (UGC), Government of India.

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Authors and Affiliations

  1. Department of Molecular Reproduction, Development, and Genetics, Indian Institute of Science, Bangalore, 560012, India

    Kartika Vashishtha, Shambhavi Shukla & Subramony Mahadevan

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  1. Kartika Vashishtha
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  2. Shambhavi Shukla
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  3. Subramony Mahadevan
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Contributions

KV and SM designed the experiments, KV and SS carried out the experiments, and KV and SM wrote the paper.

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Correspondence to Subramony Mahadevan.

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Vashishtha, K., Shukla, S. & Mahadevan, S. Involvement of BglG in Lipopolysaccharides (LPS) Synthesis and Transport in Stationary Phase in E. coli. Curr Microbiol 79, 153 (2022). https://doi.org/10.1007/s00284-022-02837-1

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