Global role for ClpP-containing proteases in stationary-phase adaptation of Escherichia coli.
To elucidate the involvement of proteolysis in the regulation of stationary-phase adaptation, the clpA, clpX, and clpP protease mutants of Escherichia coli were subjected to proteome analysis during growth and during carbon starvation. For most of the growth-phase-regulated proteins detected on our gels, the clpA, clpX, or clpP mutant failed to mount the growth-phase regulation found in the wild type. For example, in the clpP and clpA mutant cultures, the Dps protein, the WrbA protein, and the periplasmic lysine-arginine-ornithine binding protein ArgT did not display the induction typical for late-stationary-phase wild-type cells. On the other hand, in the protease mutants, a number of proteins accumulated to a higher degree than in the wild type, especially in late stationary phase. The proteins affected in this manner include the LeuA, TrxB, GdhA, GlnA, and MetK proteins and alkyl hydroperoxide reductase (AhpC). These proteins may be directly degraded by ClpAP or ClpXP, respectively, or their expression could be modulated by a protease-dependent mechanism. From our data we conclude that the levels of most major growth-phase-regulated proteins in E. coli are at some point controlled by the activity of at least one of the ClpP, ClpA, and ClpX proteins. Cultures of the strains lacking functional ClpP or ClpX also displayed a more rapid loss of viability during extended stationary phase than the wild type. Therefore, regulation by proteolysis seems to be more important, especially in resting cells, than previously suspected.
Weichart
Dieter
D
Institut für Biologie-Mikrobiologie, Freie Universitaet Berlin, Königin-Luise Strasse 12-16, 14195 Berlin, Germany. weichart@zedat.fu-berlin.de
Querfurth
Nadine
N
Dreger
Mathias
M
Hengge-Aronis
Regine
R
D000074183:ATPases Associated with Diverse Cellular Activities; D000222:Adaptation, Physiological; D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D003470:Culture Media; D049071:Endopeptidase Clp; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005947:Glucose / Q000378:metabolism; D018832:Molecular Chaperones; D009154:Mutation; D020543:Proteome; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003470:Culture Media; D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; D020543:Proteome; D012697:Serine Endopeptidases; C483568:ClpA protease, E coli; C483573:ClpXP protease, E coli; D049071:Endopeptidase Clp; D000251:Adenosine Triphosphatases; C088072:ClpX protein, E coli; D000074183:ATPases Associated with Diverse Cellular Activities; D005947:Glucose