Requirement of flhA for swarming differentiation, flagellin export, and secretion of virulence-associated proteins in Bacillus thuringiensis.
Bacillus thuringiensis is being used worldwide as a biopesticide, although increasing evidence suggests that it is emerging as an opportunistic human pathogen. While phospholipases, hemolysins, and enterotoxins are claimed to be responsible for B. thuringiensis virulence, there is no direct evidence to indicate that the flagellum-driven motility plays a role in parasite-host interactions. This report describes the characterization of a mini-Tn10 mutant of B. thuringiensis that is defective in flagellum filament assembly and in swimming and swarming motility as well as in the production of hemolysin BL and phosphatidylcholine-preferring phospholipase C. The mutant strain was determined to carry the transposon insertion in flhA, a flagellar class II gene encoding a protein of the flagellar type III export apparatus. Interestingly, the flhA mutant of B. thuringiensis synthesized flagellin but was impaired in flagellin export. Moreover, a protein similar to the anti-sigma factor FlgM that acts in regulating flagellar class III gene transcription was not detectable in B. thuringiensis, thus suggesting that the flagellar gene expression hierarchy of B. thuringiensis differs from that described for Bacillus subtilis. The flhA mutant of B. thuringiensis was also defective in the secretion of hemolysin BL and phosphatidylcholine-preferring phospholipase C, although both of these virulence factors were synthesized by the mutant. Since complementation of the mutant with a plasmid harboring the flhA gene restored swimming and swarming motility as well as secretion of toxins, the overall results indicate that motility and virulence in B. thuringiensis may be coordinately regulated by flhA, which appears to play a crucial role in the export of flagellar as well as nonflagellar proteins.
Ghelardi
Emilia
E
Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, Università degli Studi di Pisa, 56127 Pisa, Italy.
Celandroni
Francesco
F
Salvetti
Sara
S
Beecher
Douglas J
DJ
Gominet
Myriam
M
Lereclus
Didier
D
Wong
Amy C L
AC
Senesi
Sonia
S
D001413:Bacillus thuringiensis / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity* / Q000502:physiology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004251:DNA Transposable Elements; D005408:Flagellin / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006460:Hemolysin Proteins; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009068:Movement; D009154:Mutation; D015398:Signal Transduction; D014158:Transcription, Genetic; D010738:Type C Phospholipases / Q000378:metabolism; D014774:Virulence
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; C080688:FlhA protein, Bacteria; D006460:Hemolysin Proteins; D008565:Membrane Proteins; C083961:hemolysin BL protein, Bacillus; D005408:Flagellin; D010738:Type C Phospholipases; C105598:phosphatidylcholine-specific phospholipase C