ATP Regulates Calcium Efflux and Growth in E. coli
| Autor: | I. Barry Holland, Riffat Naseem, Anthony K. Campbell, Kenneth Taylor Wann |
|---|---|
| Přispěvatelé: | Institut de génétique et microbiologie [Orsay] (IGM), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2009 |
| Předmět: |
ATPase
MESH: Escherichia coli Proteins Biology MESH: Proton-Translocating ATPases MESH: Gene Expression Profiling MESH: Mutant Proteins Gene Knockout Techniques 03 medical and health sciences Polyphosphate kinase Adenosine Triphosphate Structural Biology MESH: Uncoupling Agents MESH: Adenosine Triphosphate Escherichia coli [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Molecular Biology Gene knockout MESH: Gene Knockout Techniques Oligonucleotide Array Sequence Analysis 030304 developmental biology MESH: Gene Expression Regulation Bacterial 0303 health sciences MESH: Escherichia coli Uncoupling Agents Escherichia coli Proteins Gene Expression Profiling 030302 biochemistry & molecular biology Wild type Membrane Proteins MESH: Transcription Factors Gene Expression Regulation Bacterial MESH: 2 4-Dinitrophenol Transport protein Proton-Translocating ATPases [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology Membrane protein Biochemistry MESH: Calcium MESH: Oligonucleotide Array Sequence Analysis biology.protein Calcium Mutant Proteins MESH: Membrane Proteins Efflux 2 4-Dinitrophenol Intracellular Transcription Factors |
| Zdroj: | Journal of Molecular Biology Journal of Molecular Biology, Elsevier, 2009, 391 (1), pp.42-56. ⟨10.1016/j.jmb.2009.05.064⟩ |
| ISSN: | 0022-2836 1089-8638 |
| Popis: | International audience; Escherichia coli regulates cytosolic free Ca(2+) in the micromolar range through influx and efflux. Herein, we show for the first time that ATP is essential for Ca(2+) efflux and that ATP levels also affect generation time. A transcriptome analysis identified 110 genes whose expression responded to an increase in cytosolic Ca(2+) (41 elevated, 69 depressed). Of these, 3 transport proteins and 4 membrane proteins were identified as potential Ca(2+) transport pathways. Expression of a further 943 genes was modified after 1 h in growth medium containing Ca(2+) relative to time zero. Based on the microarray results and other predicted possible Ca(2+) transporters, the level of cytosolic free Ca(2+) was measured in selected mutants from the Keio knockout collection using intracellular aequorin. In this way, we identified a knockout of atpD, coding for a component of the F(o)F(1) ATPase, as defective in Ca(2+) efflux. Seven other putative Ca(2+) transport proteins exhibited normal Ca(2+) handling. The defect in the DeltaatpD knockout cells could be explained by a 70% reduction in ATP. One millimolar glucose or 1 mM methylglyoxal raised ATP in the DeltaatpD knockout cells to that of the wild type and restored Ca(2+) efflux. One millimolar 2,4-dinitrophenol lowered the ATP in wild type to that in the DeltaatpD cells. Under these conditions, a similar defect in Ca(2+) efflux in wild type was observed in DeltaatpD cells. Ten millimolar concentration of Ca(2+) resulted in a 30% elevation in ATP in wild type and was accompanied by a 10% reduction in generation time under these conditions. Knockouts of pitB, a potential Ca(2+) transporter, atoA, the beta subunit of acetate CoA-transferase likely to be involved in polyhydroxybutyrate synthesis, and ppk, encoding polyphosphate kinase, all indicated no defect in Ca(2+) efflux. We therefore propose that ATP is most likely to regulate Ca(2+) efflux in E. coli through an ATPase. |
| Databáze: | OpenAIRE |
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