Archives of Microbiology

, Volume 198, Issue 5, pp 409–421 | Cite as

PpsA-mediated alternative pathway to complement RNase E essentiality in Escherichia coli

  • Masaru Tamura
  • Naoko Honda
  • Hirofumi Fujimoto
  • Stanley N. Cohen
  • Atsushi Kato
Original Paper

Abstract

Escherichia coli cells require RNase E, encoded by the essential gene rne, to propagate. The growth properties on different carbon sources of E. coli cells undergoing suppression of RNase E production suggested that reduction in RNase E is associated with decreased expression of phosphoenolpyruvate synthetase (PpsA), which converts pyruvate to phosphoenolpyruvate during gluconeogenesis. Western blotting and genetic complementation confirmed the role of RNase E in PpsA expression. Adventitious ppsA overexpression from a multicopy plasmid was sufficient to restore colony formation of ∆rneE. coli on minimal media containing glycerol or succinate as the sole carbon source. Complementation of ∆rne by ppsA overproduction was observed during growth on solid media but was only partial, and bacteria showed slowed cell division and grew as filamentous chains. We found that restoration of colony-forming ability by ppsA complementation occurred independent of the presence of endogenous RNase G or second-site suppressors of RNase E essentiality. Our investigations demonstrate the role of phosphoryl transfer catalyzable by PpsA as a determinant of RNase E essentiality in E. coli.

Keywords

Ribonuclease E RNase E Gluconeogenesis ppsA Phosphotransferase 

Supplementary material

203_2016_1201_MOESM1_ESM.pdf (4.2 mb)
Supplementary material 1 (PDF 4288 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Masaru Tamura
    • 1
  • Naoko Honda
    • 1
  • Hirofumi Fujimoto
    • 1
  • Stanley N. Cohen
    • 2
  • Atsushi Kato
    • 1
  1. 1.Department of Quality Assurance and Radiological ProtectionNational Institute of Infectious DiseasesTokyoJapan
  2. 2.Departments of Genetics and Medicine, School of MedicineStanford UniversityStanfordUSA

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