Document Detail

Binding sequences for RdgB, a DNA damage-responsive transcriptional activator, and temperature-dependent expression of bacteriocin and pectin lyase genes in Pectobacterium carotovorum subsp. carotovorum.
MedLine Citation:
PMID:  18689515     Owner:  NLM     Status:  MEDLINE    
Pectobacterium carotovorum subsp. carotovorum strain Er simultaneously produces the phage tail-like bacteriocin carotovoricin (Ctv) and pectin lyase (Pnl) in response to DNA-damaging agents. The regulatory protein RdgB of the Mor/C family of proteins activates transcription of pnl through binding to the promoter. However, the optimal temperature for the synthesis of Ctv (23 degrees C) differs from that for synthesis of Pnl (30 degrees C), raising the question of whether RdgB directly activates ctv transcription. Here we report that RdgB directly regulates Ctv synthesis. Gel mobility shift assays demonstrated RdgB binding to the P(0), P(1), and P(2) promoters of the ctv operons, and DNase I footprinting determined RdgB-binding sequences (RdgB boxes) on these and on the pnl promoters. The RdgB box of the pnl promoter included a perfect 7-bp inverted repeat with high binding affinity to the regulator (K(d) [dissociation constant] = 150 nM). In contrast, RdgB boxes of the ctv promoters contained an imperfect inverted repeat with two or three mismatches that consequently reduced binding affinity (K(d) = 250 to 350 nM). Transcription of the rdgB and ctv genes was about doubled at 23 degrees C compared with that at 30 degrees C. In contrast, the amount of pnl transcription tripled at 30 degrees C. Thus, the inverse synthesis of Ctv and Pnl as a function of temperature is apparently controlled at the transcriptional level, and reduced rdgB expression at 30 degrees C obviously affected transcription from the ctv promoters with low-affinity RdgB boxes. Pathogenicity toward potato tubers was reduced in an rdgB knockout mutant, suggesting that the RdgAB system contributes to the pathogenicity of this bacterium, probably by activating pnl expression.
Kazuteru Yamada; Jun Kaneko; Yoshiyuki Kamio; Yoshifumi Itoh
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Publication Detail:
Type:  Journal Article     Date:  2008-08-08
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  74     ISSN:  1098-5336     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2008 Oct 
Date Detail:
Created Date:  2008-09-26     Completed Date:  2008-10-23     Revised Date:  2013-06-05    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  6017-25     Citation Subset:  IM    
Laboratory of Applied Microbiology, Department of Microbial Biotechnology, Graduate School of Agricultural Science, Tohoku University, Tsutsumidori Amamiya-machi 1-1, Aoba-ku, Sendai 981-8555, Japan.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AB298803;  AB304880
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MeSH Terms
Bacterial Proteins / genetics*
Bacteriocins / biosynthesis*
Binding Sites
DNA Footprinting
DNA, Bacterial / chemistry,  genetics,  metabolism
Electrophoretic Mobility Shift Assay
Gene Deletion
Gene Expression Regulation, Bacterial*
Gene Order
Molecular Sequence Data
Pectobacterium carotovorum / genetics*
Plant Diseases / microbiology
Polysaccharide-Lyases / biosynthesis*
Promoter Regions, Genetic
Protein Binding
Sequence Analysis, DNA
Solanum tuberosum / microbiology
Transcription Factors / genetics*
Reg. No./Substance:
0/Bacterial Proteins; 0/Bacteriocins; 0/DNA, Bacterial; 0/Transcription Factors; 66812-91-7/carotovoricin; EC 4.2.2.-/Polysaccharide-Lyases; EC lyase

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