Mutations of the Bacillus subtilis YidC1 (SpoIIIJ) insertase alleviate stress associated with σM-dependent membrane protein overproduction

English version

Autoři: Heng Zhao ^aff001; Ankita J. Sachla ^aff001; John D. Helmann ^aff001
Působiště autorů: Department of Microbiology, Cornell University, Ithaca, NY, United States of America ^aff001
Vyšlo v časopise: Mutations of the Bacillus subtilis YidC1 (SpoIIIJ) insertase alleviate stress associated with σM-dependent membrane protein overproduction. PLoS Genet 15(10): e32767. doi:10.1371/journal.pgen.1008263
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pgen.1008263

Souhrn

In Bacillus subtilis, the extracytoplasmic function σ factor σ^M regulates cell wall synthesis and is critical for intrinsic resistance to cell wall targeting antibiotics. The anti-σ factors YhdL and YhdK form a complex that restricts the basal activity of σ^M, and the absence of YhdL leads to runaway expression of the σ^M regulon and cell death. Here, we report that this lethality can be suppressed by gain-of-function mutations in yidC1 (spoIIIJ), which encodes the major YidC membrane protein insertase in B. subtilis. B. subtilis PY79 YidC1 (SpoIIIJ) contains a single amino acid substitution in a functionally important hydrophilic groove (Q140K), and this allele suppresses the lethality of high σ^M. Analysis of a library of YidC1 variants reveals that increased charge (+2 or +3) in the hydrophilic groove can compensate for high expression of the σ^M regulon. Derepression of the σ^M regulon induces secretion stress, oxidative stress and DNA damage responses, all of which can be alleviated by the YidC1^Q140K substitution. We further show that the fitness defect caused by high σ^M activity is exacerbated in the absence of the SecDF protein translocase or σ^M-dependent induction of the Spx oxidative stress regulon. Conversely, cell growth is improved by mutation of specific σ^M-dependent promoters controlling operons encoding integral membrane proteins. Collectively, these results reveal how the σ^M regulon has evolved to up-regulate membrane-localized complexes involved in cell wall synthesis, and to simultaneously counter the resulting stresses imposed by regulon induction.

Klíčová slova:

Bacillus subtilis – Hyperexpression techniques – Insertion mutation – Integral membrane proteins – Membrane proteins – Point mutation – Polymerase chain reaction – Regulons

Zdroje

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