title: PpsA-mediated alternative pathway to complement RNase E essentiality in Escherichia coli.
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 ∆rne E. 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.
authors.affiliation: Department of Quality Assurance and Radiological Protection, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan. mtamura@niid.go.jp.
authors.forename: Masaru
authors.initials: M
authors.lastname: Tamura
authors.affiliation: Department of Quality Assurance and Radiological Protection, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Honda
authors.affiliation: Department of Quality Assurance and Radiological Protection, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Fujimoto
authors.affiliation: Departments of Genetics and Medicine, School of Medicine, Stanford University, Stanford, CA, USA.
authors.forename: Stanley N
authors.initials: SN
authors.lastname: Cohen
authors.affiliation: Department of Quality Assurance and Radiological Protection, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kato
mesh_terms: D004722:Endoribonucleases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D010957:Plasmids / Q000235:genetics; D050743:Pyruvate Synthase / Q000235:genetics / Q000378:metabolism*; D019289:Pyruvic Acid / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D019289:Pyruvic Acid; D050743:Pyruvate Synthase; C506224:ppsA protein, E coli; D004722:Endoribonucleases; C022537:ribonuclease E
keywords: Gluconeogenesis; Phosphotransferase; RNase E; Ribonuclease E; ppsA