Article

RelA-dependent (p)ppGpp production controls exoenzyme synthesis in Erwinia carotovora subsp. atroseptica.

Department of Biochemistry, Tennis Court Road, University of Cambridge, Cambridge CB2 1QW, United Kingdom.
Journal of Bacteriology (impact factor: 3.83). 12/2007; 189(21):7643-52. DOI:10.1128/JB.00920-07 pp.7643-52
Source: PubMed

ABSTRACT In this report, we investigate the link between nutrient limitation, RelA-mediated (p)ppGpp production, and virulence in the phytopathogen Erwinia carotovora subsp. atroseptica. A relA null mutant (JWC7) was constructed by allelic exchange, and we confirmed that, unlike the wild-type progenitor, this mutant did not produce elevated levels of (p)ppGpp upon nutrient downshift. However, (p)ppGpp production could be restored in strain JWC7 during nutrient limitation by supplying relA in trans. During growth on exoenzyme-inducing minimal medium, the relA mutant showed a diminution in secreted pectate lyase and protease activities and a severe defect in motility. The relA mutant was also impaired in its ability to cause rot in potato tubers. In the presence of serine hydroxamate (a competitive inhibitor of seryl tRNA synthase and a potent inducer of the stringent response in wild-type E. carotovora subsp. atroseptica), exoenzyme production was essentially abolished in JWC7 but could be restored in the presence of plasmid-borne relA. The inhibition of exoenzyme production in JWC7 caused by serine hydroxamate could not be overcome by addition of the quorum-sensing signal molecule, N-3-oxohexanoyl-l-homoserine lactone. Quantitative reverse transcription-PCR analysis of selected RNA species confirmed that the effects of relA on secreted pectate lyase activity and motility could be attributed to a reduction in transcription of the corresponding genes. We conclude that nutrient limitation is a potent environmental cue that triggers (p)ppGpp-dependent exoenzyme production in E. carotovora subsp. atroseptica. Furthermore, our data suggest that nutrient limitation [or rather, (p)ppGpp accumulation] is a prerequisite for effective quorum-sensing-dependent activation of exoenzyme production.

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Keywords

cause rot
 
E. carotovora subsp
 
effective quorum-sensing-dependent activation
 
exoenzyme production
 
exoenzyme-inducing minimal medium
 
nutrient limitation [or
 
p)ppGpp accumulation]
 
p)ppGpp production
 
p)ppGpp-dependent exoenzyme production
 
phytopathogen Erwinia carotovora subsp
 
plasmid-borne relA
 
potent environmental cue
 
quorum-sensing signal molecule
 
relA null mutant
 
RNA species
 
secreted pectate lyase
 
secreted pectate lyase activity
 
serine hydroxamate
 
seryl tRNA synthase
 
wild-type E. carotovora subsp