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Using Microtiter Dish Radiolabeling for Multiple In Vivo Measurements Of Escherichia coli pppGpp Followed by Thin Layer Chromatography
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Using Microtiter Dish Radiolabeling for Multiple In Vivo Measurements Of Escherichia coli pppGpp Followed by Thin Layer Chromatography

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Effect of the RNA pyrophosphohydrolase RppH on envelope integrity in Escherichia coli

Umji Choi1, Young-Ha Park2, Yeon-Ran Kim2

  • 1Department of Biological Sciences, Myongji University, San 38-2, Namdong, Yongin, Gyeonggido 449-728, Republic of Korea.

FEMS Microbiology Letters
|September 2, 2017
PubMed

Insights

The bacterial enzyme RppH regulates mRNA decay. Loss of RppH in Escherichia coli causes sensitivity to antibiotics and envelope stresses, revealing its role in maintaining cell envelope integrity.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • The bacterial enzyme RppH initiates mRNA decay by removing pyrophosphate from 5'-triphosphorylated mRNA.
  • While Escherichia coli RppH has promiscuous substrate specificity, few transcripts are affected by its loss.
  • The physiological role of RppH necessitates phenotypic analysis of the rppH mutant, which has not been previously determined.

Purpose of the Study:

  • To determine the phenotypes of the rppH mutant.
  • To understand the physiological role of RppH in Escherichia coli.
  • To investigate the association of RppH with bacterial envelope integrity.

Main Methods:

  • Phenotypic analysis of the rppH mutant.
  • Drug susceptibility testing.
  • Envelope stress testing (osmotic stress, ethanol, sodium dodecyl sulfate).

Main Results:

  • The rppH mutant exhibited sensitivity to various chemicals, including antibiotics and envelope stresses.
  • All observed phenotypes were attributed to the loss of RppH enzymatic activity.
  • The rppH mutant displayed increased envelope permeability, while increased RppH activity inhibited growth at low temperatures.

Conclusions:

  • The study identified several phenotypes of the rppH mutant linked to envelope integrity.
  • RppH plays a crucial role in maintaining bacterial cell envelope integrity.
  • Loss of RppH function compromises envelope integrity, leading to sensitivity to various stresses.
Keywords:
RNA pyrophosphohydrolaseantibiotic sensitivitycold stress resistanceenvelope integrityenvelope stress

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