PMID:14766922
| Citation |
Kuo, MS, Chen, KP and Wu, WF (2004) Regulation of RcsA by the ClpYQ (HslUV) protease in Escherichia coli. Microbiology (Reading, Engl.) 150:437-46 |
|---|---|
| Abstract |
Escherichia coli ClpYQ protease and Lon protease possess a redundant function for degradation of SulA, a cell division inhibitor. An experimental cue implied that the capsule synthesis activator RcsA, a known substrate of Lon, is probably a specific substrate for the ClpYQ protease. This paper shows that overexpression of ClpQ and ClpY suppresses the mucoid phenotype of a lon mutant. Since the cpsB (wcaB) gene, involved in capsule synthesis, is activated by RcsA, the reporter construct cpsB-lacZ was used to assay for beta-galactosidase activity and thus follow RcsA stability. The expression of cpsB-lacZ was increased in double mutants of lon in combination with clpQ or/and clpY mutation(s) compared with the wild-type or lon single mutants. Overproduction of ClpYQ or ClpQ decreased cpsB-lacZ expression. Additionally, a P(BAD)-rcsA fusion construct showed quantitatively that an inducible RcsA activates cpsB-lacZ expression. The effect of RcsA on cpsB-lacZ expression was shown to be influenced by the ClpYQ activities. Moreover, a rcsA(Red)-lacZ translational fusion construct showed higher activity of RcsA(Red)-LacZ in a clpQ clpY strain than in the wild-type. By contrast, overproduction of cellular ClpYQ resulted in decreased beta-galactosidase levels of RcsA(Red)-LacZ. Taken together, the data indicate that ClpYQ acts as a secondary protease in degrading the Lon substrate RcsA. |
| Links | |
| Keywords |
ATP-Dependent Proteases; Base Sequence; DNA Primers; Endopeptidase Clp; Escherichia coli Proteins/genetics; Gene Expression Regulation, Bacterial/genetics; Gene Expression Regulation, Enzymologic/genetics; Genotype; Heat-Shock Proteins/genetics; Heat-Shock Proteins/metabolism; Kinetics; Polymerase Chain Reaction; Protease La; Recombinant Fusion Proteins/metabolism; Serine Endopeptidases/genetics; Serine Endopeptidases/metabolism |
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