FliG and FliM distribution in the Salmonella typhimurium cell and flagellar basal bodies.
Salmonella typhimurium FliG and FliM are two of three proteins known to be necessary for flagellar morphogenesis as well as energization and switching of flagellar rotation. We have determined FliG and FliM levels in cellular fractions and in extended flagellar basal bodies, using antibodies raised against the purified proteins. Both proteins were found predominantly in the detergent-solubilized particulate fraction containing flagellar structures. Basal flagellar fragments could be separated from partially constructed basal bodies by gel filtration chromatography. FliG and FliM were present in an approximately equimolar ration in all gel-filtered fractions. FliG and FliM copy numbers, estimated relative to that of the hook protein from the early fractions containing long, basal, flagellar fragments, were (means +/- standard errors) 41 +/- 10 and 37 +/- 13 per flagellum, respectively. Extended structures were present in the earliest identifiable basal bodies. Immunoelectron microscopy and immunoblot gel analysis suggested that the FliG and, to a less certain degree, the FliM contents of these structures were the same as those for the complete basal bodies. These facts are consistent with the postulate that FliG and FliM affect flagellar morphogenesis as part of the extended basal structure, formation of which is necessary for assembly of more-distal components of the flagellum. The determined stoichiometries will provide important constraints to modelling energization and switching of flagellar rotation.
Department of Physiology & Biophysics, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
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Zhao
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Amsler
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Matsumura
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Khan
D001426:Bacterial Proteins / Q000032:analysis*; D002458:Cell Fractionation / Q000379:methods; D002850:Chromatography, Gel; D005407:Flagella / Q000737:chemistry* / Q000648:ultrastructure; D012486:Salmonella typhimurium / Q000737:chemistry* / Q000648:ultrastructure; D014461:Ultracentrifugation
D001426:Bacterial Proteins; C075660:Flig protein, Bacteria; C066821:FliM protein, Bacteria