FlhF, a signal recognition particle-like GTPase, is involved in the regulation of flagellar arrangement, motility behaviour and protein secretion in Bacillus cereus.
Flagellar arrangement is a highly conserved feature within bacterial species. However, only a few genes regulating cell flagellation have been described in polar flagellate bacteria. This report demonstrates that the arrangement of flagella in the peritrichous flagellate Bacillus cereus is controlled by flhF. Disruption of flhF in B. cereus led to a reduction in the number of flagella from 10-12 to 1-3 filaments per cell in the insertion mutant MP06. Moreover, compared to the parental strain, MP06 exhibited: (i) shorter smooth swimming phases, causing reduced swimming motility but not affecting chemotaxis; (ii) complete inhibition of swarming motility, as differentiated swarm cells were never detected; (iii) an increased amount of extracellular proteins; and (iv) differential export of virulence determinants, such as haemolysin BL (HBL), phosphatidylcholine-preferring phospholipase C (PC-PLC) and non-haemolytic enterotoxin (NHE). Introduction of a plasmid harbouring flhF (pDGflhF) into MP06 completely restored the wild-type phenotype in the trans-complemented strain MP07. B. cereus flhF was found to constitute a monocistronic transcriptional unit and its overexpression did not produce abnormal features in the wild-type background. Characterization of a B. cereus mutant (MP05) carrying a partial flhF deletion indicated that the last C-terminal domain of FlhF is involved in protein export while not required for flagellar arrangement and motility behaviour. Taken together, these data suggest that B. cereus FlhF is a promising candidate for connecting diverse cellular functions, such as flagellar arrangement, motility behaviour, pattern of protein secretion and virulence phenotype.
Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, Università di Pisa, via San Zeno 37, 56127 Pisa, Italy.
Sara
S
Salvetti
Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, Università di Pisa, via San Zeno 37, 56127 Pisa, Italy.
Emilia
E
Ghelardi
Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, Università di Pisa, via San Zeno 37, 56127 Pisa, Italy.
Francesco
F
Celandroni
Dipartimento di Biologia, Sezione di Microbiologia, Università di Pisa, via San Zeno 35, 56127 Pisa, Italy.
Mara
M
Ceragioli
Dipartimento di Morfologia Umana e Biologia Applicata, Università di Pisa, via Roma 55, 56126 Pisa, Italy.
Francesco
F
Giannessi
Dipartimento di Biologia, Sezione di Microbiologia, Università di Pisa, via San Zeno 35, 56127 Pisa, Italy.
Sonia
S
Senesi
D001409:Bacillus cereus / Q000235:genetics / Q000502:physiology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005407:Flagella / Q000235:genetics / Q000502:physiology*; D017353:Gene Deletion; D005796:Genes; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D020559:Monomeric GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D009068:Movement / Q000502:physiology; D016254:Mutagenesis, Insertional; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D017422:Sequence Analysis, DNA; D037521:Virulence Factors / Q000378:metabolism
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D037521:Virulence Factors; C078317:flhF protein, Bacteria; D020559:Monomeric GTP-Binding Proteins