Downregulation of the motA gene delays the escape of the obligate predator Bdellovibrio bacteriovorus 109J from bdelloplasts of bacterial prey cells.
Bdellovibrio bacteriovorus is a Gram-negative bacterium that preys on other Gram-negative bacteria. The lifecycle of B. bacteriovorus alternates between an extracellular flagellated and highly motile non-replicative attack-phase cell and a periplasmic non-flagellated growth-phase cell. The prey bacterium containing periplasmic bdellovibrios becomes spherical but osmotically stable, forming a structure known as the bdelloplast. After completing the growth phase, newly formed bdellovibrios regain their flagellum and escape the bdelloplast into the environment, where they encounter more prey bacteria. The obligate predatory nature of B. bacteriovorus imposes a major difficulty to introducing mutations in genes directly involved in predation, since these mutants could be non-viable. This work reports the cloning of the B. bacteriovorus 109J motAB operon, encoding proteins from the flagellar motor complex, and a genetic approach based on the expression of a motA antisense RNA fragment to downregulate motility. Periplasmic bdellovibrios carrying the plasmid expressing antisense RNA displayed a marked delay in escaping from bdelloplasts, while the released attack-phase cells showed altered motility. These observations suggest that a functionally intact flagellar motor is required for the predatory lifecycle of B. bacteriovorus. Also, the use of antisense RNA expression may be a useful genetic tool to study the Bdellovibrio developmental cycle.
Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada N6A 5C1.
Ronald S
RS
Flannagan
Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada N6A 5C1.
Miguel A
MA
Valvano
Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada N6A 5C1.
Susan F
SF
Koval
D001426:Bacterial Proteins / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D001501:Bdellovibrio / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity*; D004269:DNA, Bacterial / Q000235:genetics; D015536:Down-Regulation; D005798:Genes, Bacterial; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon; D010957:Plasmids / Q000235:genetics; D016372:RNA, Antisense / Q000235:genetics; D012329:RNA, Bacterial / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008164:Luminescent Proteins; C063010:MotA protein, Bacteria; D016372:RNA, Antisense; D012329:RNA, Bacterial; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins