title: Nutrient dependence of RNase E essentiality in Escherichia coli.
abstract: Escherichia coli cells normally require RNase E activity to form colonies (colony-forming ability [CFA]). The CFA-defective phenotype of cells lacking RNase E is partly reversed by overexpression of the related endoribonuclease RNase G or by mutation of the gene encoding the RNA helicase DeaD. We found that the carbon source utilization by rne deaD doubly mutant bacteria differs from that of rne(+) cells and from that of cells mutated in deaD alone and that the loss of rne function in these bacteria limits conversion of the glycolytic pathway product phosphoenolpyruvate to the tricarboxylic acid (TCA) cycle intermediate oxaloacetic acid. We show that the mechanism underlying this effect is reduced production of the enzyme phosphoenolpyruvate carboxylase (PPC) and that adventitious overexpression of PPC, which facilitates phosphoenolpyruvate utilization and connects the glycolytic pathway with the TCA cycle, restored CFA to rne deaD mutant bacteria cultured on carbon sources that otherwise were unable to sustain growth. We further show that bacteria producing full-length RNase E, which allows formation of degradosomes, have nutritional requirements different from those of cells supplied with only the N-terminal catalytic region of RNase E and that mitigation of RNase E deficiency by overexpression of a related RNase, RNase G, is also affected by carbon source. Our results reveal previously unsuspected effects of RNase E deficiency and degradosome formation on nutrient utilization by E. coli cells.
authors.affiliation: Department of Genetics, School of Medicine, Stanford University, Stanford, California, USA.
authors.forename: Masaru
authors.initials: M
authors.lastname: Tamura
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Moore
authors.forename: Stanley N
authors.initials: SN
authors.lastname: Cohen
mesh_terms: D002244:Carbon / Q000378:metabolism; D002952:Citric Acid Cycle / Q000235:genetics; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism*; D006019:Glycolysis / Q000235:genetics; D006801:Humans; D062907:Oxaloacetic Acid / Q000378:metabolism; D010728:Phosphoenolpyruvate / Q000378:metabolism; D010730:Phosphoenolpyruvate Carboxylase / Q000378:metabolism; D020365:RNA Helicases / Q000235:genetics / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; D062907:Oxaloacetic Acid; D010728:Phosphoenolpyruvate; D002244:Carbon; D004722:Endoribonucleases; C081905:rng protein, E coli; C022537:ribonuclease E; D020365:RNA Helicases; D010730:Phosphoenolpyruvate Carboxylase