The dsbA genes, which encode major periplasmic disulfide-bond-forming proteins, were isolated from subsp. (Ecc) and subsp. (Eca), and the , encoding another periplasmic disulfide oxidoreductase was isolated from Ecc. All three genes were sequenced and mutants deficient in these genes were created by marker exchange mutagenesis. The Ecc mutants were severely affected in activity and secretion of pectate lyase, probably due to the absence of functional PelC, which is predicted to require disulfide bond formation to achieve its correct conformation prior to secretion across the outer membrane. Similarly, endopolygalacturonase, also predicted to possess disulfide bonds, displayed reduced activity. The major Ecc cellulase (CeIV) does not contain cysteine residues and was still secreted in -deficient strains. This observation demonstrated unequivocally that the localization and activity of the individual components of the Out apparatus are independent of disulfide bond formation. Surprisingly, cellulase activity was shown to be increased ~ two- to threefold in the DsbA mutant. This phenomenon resulted from transcriptional up-regulation of cel/V gene expression. In contrast, transcription of both pe/C and were down-regulated in -deficient strains when compared to the wild-type. Protease (Prt) activity and secretion were unaffected in the mutant. Prt activity was considerably reduced in the double mutant. However Prt was secreted normally in this strain. The mutant was found to be non-motile, suggesting that disulfide bond formation is essential for motility in this strain. All of the mutants showed reduced tissue maceration These results suggest that a feedback regulation system operates in Ecc. In this system, defects in periplasmic disulfide bond formation act as a signal which is relayed to the transcription machinery regulating gene expression in diverse ways.


Article metrics loading...

Loading full text...

Full text loading...


Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error