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1.
BMC Genomics ; 17(1): 914, 2016 11 14.
Article in English | MEDLINE | ID: mdl-27842489

ABSTRACT

BACKGROUND: In a process known as phase variation, the marine bacterium and cholera pathogen Vibrio cholerae alternately expresses smooth or rugose colonial phenotypes, the latter being associated with advanced biofilm architecture and greater resistance to ecological stress. To define phase variation at the transcriptomic level in pandemic V. cholerae O1 El Tor strain N16961, we compared the RNA-seq-derived transcriptomes among the smooth parent N16961, its rugose derivative (N16961R) and a smooth form obtained directly from the rugose at high frequencies consistent with phase variation (N16961SD). RESULTS: Differentially regulated genes which clustered into co-expression groups were identified for specific cellular functions, including acetate metabolism, gluconeogenesis, and anaerobic respiration, suggesting an important link between these processes and biofilm formation in this species. Principal component analysis separated the transcriptome of N16961SD from the other phase variants. Although N16961SD was defective in biofilm formation, transcription of its biofilm-related vps and rbm gene clusters was nevertheless elevated as judged by both RNA-seq and RT-qPCR analyses. This transcriptome signature was shared with N16961R, as were others involving two-component signal transduction, chemotaxis, and c-di-GMP synthesis functions. CONCLUSIONS: Precise turnarounds in gene expression did not accompany reversible phase transitions (i.e., smooth to rugose to smooth) in the cholera pathogen. Transcriptomic signatures consisting of up-regulated genes involved in biofilm formation, environmental sensing and persistence, chemotaxis, and signal transduction, which were shared by N16961R and N16961SD variants, may implicate a stress adaptation in the pathogen that facilitates transition of the N16961SD smooth form back to rugosity should environmental conditions dictate.


Subject(s)
Adaptation, Biological/genetics , Cholera/microbiology , Stress, Physiological/genetics , Transcriptome , Vibrio cholerae/genetics , Acetates/metabolism , Biofilms , Biological Transport , Carbohydrate Metabolism , Gene Expression Profiling , Gene Expression Regulation, Bacterial , High-Throughput Nucleotide Sequencing , Phenotype , Protein Transport , Vesicular Transport Proteins/genetics , Vesicular Transport Proteins/metabolism , Vibrio cholerae/metabolism
2.
FEBS Lett ; 590(24): 4564-4572, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27859050

ABSTRACT

The human pathogen Vibrio vulnificus undergoes phase variation among colonial morphotypes, including a virulent opaque form which produces capsular polysaccharide (CPS) and a translucent phenotype that produces little or no CPS and is attenuated. Here, we found that a V. vulnificus mutant defective for RfaH antitermination control showed a diminished capacity to undergo phase variation and displayed significantly reduced distal gene expression within the Group I CPS operon. Moreover, the rfaH mutant produced negligible CPS and was highly sensitive to killing by normal human serum, results which indicate that RfaH is likely essential for virulence in this bacterium.


Subject(s)
Bacterial Capsules/metabolism , Bacterial Proteins/genetics , Gene Expression Regulation, Bacterial , Peptide Elongation Factors/genetics , Polysaccharides, Bacterial/biosynthesis , Vibrio vulnificus/metabolism , Virulence Factors/genetics , Bacterial Capsules/drug effects , Bacterial Capsules/genetics , Bacterial Proteins/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Humans , Immunoglobulins/blood , Immunoglobulins/pharmacology , Microbial Viability/drug effects , Mutation , Operon , Peptide Elongation Factors/deficiency , Phenotype , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Trans-Activators/deficiency , Trans-Activators/genetics , Vibrio vulnificus/drug effects , Vibrio vulnificus/genetics , Vibrio vulnificus/pathogenicity , Virulence Factors/deficiency
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