A gene cluster for amylovoran synthesis in Erwinia amylovora: characterization and relationship to cps genes in Erwinia stewartii

1993 ◽  
Vol 239 (1-2) ◽  
pp. 158-168 ◽  
Author(s):  
Frank Bernhard ◽  
David L. Coplin ◽  
Klaus Geider
Keyword(s):  
Gene Cluster ◽  
Erwinia Amylovora ◽  
Erwinia Stewartii

Characterization of a gene locus from Erwinia amylovora with regulatory functions in exopolysaccharide synthesis of Erwinia spp.

1998 ◽  
Vol 44 (7) ◽  
pp. 657-666 ◽  
Author(s):  
Phillip Aldridge ◽  
Frank Bernhard ◽  
Peter Bugert ◽  
David L Coplin ◽  
Klaus Geider
Keyword(s):  
Structural Gene ◽  
Erwinia Amylovora ◽  
Genomic Library ◽  
Expression Vectors ◽  
External Signal ◽  
Wild Type ◽  
Reading Frame ◽  
Gus Reporter ◽  
Acid Protein ◽  
Erwinia Stewartii

In a genomic library of Erwinia amylovora, a locus has been identified that can suppress an Erwinia stewartii rcsA mutant. In addition, the locus induced a mucoid sticky phenotype of colonies in a wild-type strain of Erwinia stewartii and increased exopolysaccharide synthesis in several species of bacteria belonging to the genus Erwinia. An open reading frame was identified at this locus encoding a 225 amino acid protein that contained a helix-turn-helix motif typical of transcriptional regulators. The corresponding gene was subsequently named rcsV (regulator of capsular synthesis affecting viscosity). A mutant of rcsV in wild-type Erwinia amylovora had no detectable phenotype and produced typical levels of amylovoran under laboratory conditions. The rcsV gene on a high copy number plasmid under the control of its own promoter did not alter amylovoran production, in contrast to in-frame fusions of the structural gene in expression vectors. Since even the lac promoter was inert in the expression of rcsV, a DNA-binding protein could inhibit transcription of the gene in Erwinia amylovora. On the other hand, an Erwinia amylovora rcsA mutant was suppressed by rcsV when its promoter was replaced and the structural gene fused in-frame with lacZ' or malE. Northern blots, with total RNA from Erwinia amylovora, or promoter analysis using the GUS reporter gene did not show expression of rcsV in Erwinia amylovora, although primer extension analysis did. RcsV could be a component involved in the regulation of amylovoran synthesis, and gene expression may require an unknown external signal during the life cycle or pathogenesis of Erwinia amylovora. Key words: amylovoran, fire blight, rcsA-like activator, fusion protein.

GENETICS AND COMPLEMENTATION OF DNA REGIONS INVOLVED IN AMYLOVORAN SYNTHESIS OF ERWINIA AMYLOVORA AND STEWARTAN SYNTHESIS OF ERWINIA STEWARTII

1996 ◽  
pp. 269-274 ◽  
Author(s):  
F. Bernhard ◽  
D. Schullerus ◽  
P. Bellemann ◽  
K. Geider ◽  
D.R. Majerczak ◽  
...  
Keyword(s):  
Erwinia Amylovora ◽  
Erwinia Stewartii

scholarly journals Genetic Organization of the hrp Gene Cluster and dspAE/BF Operon in Erwinia herbicola pv. gypsophilae

2001 ◽  
Vol 14 (3) ◽  
pp. 431-436 ◽  
Author(s):  
Henia Mor ◽  
Shulamit Manulis ◽  
Michal Zuck ◽  
Roni Nizan ◽  
David L. Coplin ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Erwinia Amylovora ◽  
Amino Acid Identity ◽  
Specific Region ◽  
Gall Formation ◽  
Erwinia Herbicola ◽  
Acid Identity ◽  
Pantoea Stewartii ◽  
Hrp Genes ◽  
Disease Specific

Erwinia herbicola pv. gypsophilae induces gall formation in gypsophila that is dependent on the existence of a pathogenicity plasmid (pPATHEhg). We previously demonstrated the presence of several hrp genes on this plasmid. By employing transposon mutagenesis and sequencing, a functional hrp gene cluster on the pPATHEhg has now been characterized completely. The hrp genes of E. herbicola pv. gypsophilae are remarkably similar to and colinear with those of Erwinia amylovora and Pantoea stewartii and generally showed 60 to 90% nucleotide or deduced amino acid identity. E. herbicola pv. gypsophilae, however, lacks hrpW, which is present in E. amylovora. Additionally, E. herbicola pv. gypsophilae mutants deficient in harpin production retained pathogenicity and were slightly reduced in their ability to elicit a hypersensitive response (HR) in tobacco. The “disease specific ” region, dspA/EB/F, exhibited 60 to 74% identity with the dspA/EB/F loci of E. amylovora and P. stewartii, respectively. Mutations in dspA/E abolished pathogenicity of E. herbicola pv. gypsophilae but not HR elicitation on tobacco. Inactivation of HrpL reduced plant-induced transcription of dspA/E by three orders, indicating Hrp-dependent regulation.

scholarly journals Role of Exopolysaccharides in the Survival and Pathogenesis of the Fire Blight Bacterium, Erwinia amylovora

1994 ◽  
Author(s):  
David Gutnick ◽  
David L. Coplin
Keyword(s):  
Escherichia Coli ◽  
Pathogenic Bacteria ◽  
Erwinia Amylovora ◽  
Extracellular Polysaccharide ◽  
Gene Clusters ◽  
Site Directed Mutagenesis ◽  
E Coli ◽  
Erwinia Stewartii ◽  
Stewart's Wilt

Fireblight, a disease of apples and pears, is caused by Erwinia amylovora. Mutants of E. amylovora that do not produce the extreacellular polysaccharide (EPS), amylovoran, are avirulent. A similar EPS, stewartan, is produced by E. stewartii, which caused Stewart's wilt of corn, and which has also been implicated in the virulence of this strain. Both stewartan and amylovoran are type 1 capsular polysaccharides, typified by the colanic acid slime produced by Escherichia coli. Extracellular polysaccharide slime and capsules are important for the virulence of bacterial pathogens of plants and animals and to enhance their survival and dissemination outside of the host. The goals of this project were to examine the importance of polysaccharide structure on the pathogenicity and survival properties of three pathogenic bacteria: Erwinia amylovora, Erwinia stewartii and Escherichia coli. The project was a collaboration between the laboratories of Dr. Gutnick (PI, E. coli genetics and biochemistry), Dr. Coplin (co-PI, E. stewartii genetics) and Dr. Geider (unfunded collaborator, E. amylovora genetics and EPS analysis). Structural analysis of the EPSs, sequence analysis of the biosynthetic gene clusters and site-directed mutagenesis of individual cps and ams genes revealed that the three gene clusters shared common features for polysaccharide polymerization, translocation, and precursor synthesis as well as in the modes of transcriptional regulation. Early EPS production resulted in decreased virulence, indicating that EPS, although required for pathogenicity, is anot always advantageous and pathogens must regulate its production carefully.

The hrp Gene Cluster of Erwinia Amylovora

1991 ◽  
pp. 53-60 ◽  
Author(s):  
S. V. Beer ◽  
D. W. Bauer ◽  
X. H. Jiang ◽  
R. J. Laby ◽  
B. J. Sneath ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Erwinia Amylovora

scholarly journals Fe2+ chelator proferrorosamine A: a gene cluster of Erwinia rhapontici P45 involved in its synthesis and its impact on growth of Erwinia amylovora CFBP1430

Microbiology ◽  
2016 ◽  
Vol 162 (2) ◽  
pp. 236-245 ◽  
Author(s):  
Yannick Born ◽  
Mitja N. P. Remus-Emsermann ◽  
Marco Bieri ◽  
Tim Kamber ◽  
Jörn Piel ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Erwinia Amylovora ◽  
Erwinia Rhapontici
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