Sensing structural intermediates in bacterial flagellar assembly.

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Sensing structural intermediates in bacterial
flagellar assembly. Hughes et al., 1993 Science
262 1277-1280
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Building a flagellum
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EnvZ kinase OmpRP
Glucose Osmolarity Heat Shock
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FljB Filament Fla- HBB- FliA sigma 28
Relative levels of FlgM and FliA are important
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Is FlgM shipped out?
WT
M-
Hk-
A-
Fil-
Cap-
2- 1-
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Overexpressing FliA from a plasmid
WT
M- 2- 1- A- Fil- Cap- Hk-
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Behavior of FlgM mutants on motility plates
Right FlgM-Lac Right high
FlgM Motile revertants
White? Blue?
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Cap-
M-Z tnsl
M-Z tnscrp
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Conclusions
1. Relative levels of FlgM and FliA are
important 2. FlgM is secreted outside this
requires a complete HBB 3. Attaching a bulky
protein to FlgM interfers with FlgM secretion
as evidenced by lack of motility and lack of
external flagellin
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EnvZ kinase OmpRP
Glucose Osmolarity Heat Shock
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Testing regulation of flagellar length model
Flagella stained with Texas Red-conjugated
antibody
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Testing quick response model
After shearing
Flagellate non-flagellate controls
Rosu Hughes, J. Bact. 2006
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FlgM secretion after shearing (blending)
IC Intracellular EC - Extracellular
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De-flagellation did not affect class 3 fliC
transcription
fliC-lacZ transcription
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FlgM is a drought sensor