Document Detail


Molecular characterization and expression of the Erwinia carotovora hrpNEcc gene, which encodes an elicitor of the hypersensitive reaction.
MedLine Citation:
PMID:  9150595     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The nucleotide sequence of hrpNEcc DNA, cloned from Erwinia carotovora subsp. carotovora strain Ecc71, reveals a coding region of 1,068 bp which matches the size of hrpNEcc transcripts. hrpNEcc is predicted to encode a glycine-rich protein of approximately 36 kDa. Like the elicitors of the hypersensitive reaction (HR) produced by E. chrysanthemi (HarpinEch) and E. amylovora (HarpinEa), the deduced 36-kDa protein does not possess a typical signal sequence, but it contains a putative membrane-spanning domain. In Escherichia coli strains overexpressing hrpNEcc, the 36-kDa protein has been identified as the hrpNEcc product by Western blot analysis using anti-HarpinEch antibodies. The 36-kDa protein fractionated from E. coli elicits the HR in tobacco leaves. Moreover, a HrpN- and RsmA- double mutant (RsmA = regulator of secondary metabolites) does not produce this 36-kDa protein or elicit the HR, although this strain, like the RsmA- and HrpN+ bacteria, overproduces extracellular enzymes and macerates celery petioles. These observations demonstrate that hrpNEcc encodes the elicitor of the HR, designated HarpinEcc. The levels of hrpNEcc transcripts are affected in both RsmA+ and RsmA- strains by media composition and carbon sources, although the mRNA levels are substantially higher in the RsmA- strains. The expression of hrpNEcc in Ecc71 is cell density dependent and is activated by the quorum-sensing signal, N-(3-oxohexanoyl)-L-homoserine lactone (OHL). By contrast, hrpNEcc expression in an RsmA- strain is independent of cell density, and substantial expression occurs in the absence of OHL. The effects of cultural conditions and the occurrence of putative cis-acting sequences, such as consensus sigma 54 promoters and an hrp promoter upstream of the transcriptional start site, indicate that the production of HarpinEcc in wild-type RsmA+ E. carotovora subsp. carotovora is tightly regulated. These observations, taken along with the finding that the HR is caused by RsmA- mutants but not by RsmA+ strains (Cui et al., 1996, Mol. Plant-Microbe Interact. 9:565-573), strongly support the idea that the inability of the wild-type pectolytic E. carotovora subsp. carotovora to elicit the HR is due to the lack of a significant level of HarpinEcc production.
Authors:
A Mukherjee; Y Cui; Y Liu; A K Chatterjee
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Molecular plant-microbe interactions : MPMI     Volume:  10     ISSN:  0894-0282     ISO Abbreviation:  Mol. Plant Microbe Interact.     Publication Date:  1997 May 
Date Detail:
Created Date:  1997-06-26     Completed Date:  1997-06-26     Revised Date:  2011-01-07    
Medline Journal Info:
Nlm Unique ID:  9107902     Medline TA:  Mol Plant Microbe Interact     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  462-71     Citation Subset:  IM    
Affiliation:
Department of Plant Pathology, University of Missouri, Columbia 65211, USA.
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MeSH Terms
Descriptor/Qualifier:
4-Butyrolactone / analogs & derivatives,  metabolism
Amino Acid Sequence
Bacterial Outer Membrane Proteins / biosynthesis,  genetics*
Bacterial Proteins / metabolism
Base Sequence
Culture Media
Erwinia / genetics*,  pathogenicity
Gene Expression Regulation, Bacterial*
Genes, Bacterial*
Molecular Sequence Data
Plant Diseases / microbiology
Plants, Toxic
RNA-Binding Proteins*
Recombinant Proteins / biosynthesis
Repressor Proteins / metabolism
Sequence Homology, Amino Acid
Tobacco / microbiology
Transcription, Genetic
Vegetables / microbiology
Chemical
Reg. No./Substance:
0/Bacterial Outer Membrane Proteins; 0/Bacterial Proteins; 0/Culture Media; 0/RNA-Binding Proteins; 0/Recombinant Proteins; 0/Repressor Proteins; 0/RsmA protein, Erwinia carotovora; 0/harpin protein, Erwinia amylovora; 151217-41-3/HrpZ protein, Pseudomonas syringae; 76924-95-3/N-(3-oxohexanoyl)-3-aminodihydro-2(3H)-furanone; 96-48-0/4-Butyrolactone

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