9 results on '"Aizawa, Shin-Ichi"'
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2. Length of the Flagellar Hook and the Capacity of the Type III Export Apparatus
- Author
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Makishima, Shigeru, Komoriya, Kaoru, Yamaguchi, Shigeru, and Aizawa, Shin-Ichi
- Published
- 2001
3. The role of the FliK molecular ruler in hook-length control in Salmonella enterica.
- Author
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Erhardt, Marc, Hirano, Takanori, Su, Yichu, Paul, Koushik, Wee, Daniel H., Mizuno, Shino, Aizawa, Shin-Ichi, and Hughes, Kelly T.
- Subjects
CATALYSIS ,PROTEINS ,MOLECULES ,FLAGELLATA ,GENETIC mutation ,SECRETION - Abstract
A molecular ruler, FliK, controls the length of the flagellar hook. FliK measures hook length and catalyses the secretion-substrate specificity switch from rod-hook substrate specificity to late substrate secretion, which includes the filament subunits. Here, we show normal hook-length control and filament assembly in the complete absence of the C-ring thus refuting the previous ‘cup’ model for hook-length control. Mutants of C-ring components, which are reported to produce short hooks, show a reduced rate of hook–basal body assembly thereby allowing for a premature secretion-substrate specificity switch. Unlike fliK null mutants, hook-length control in an autocleavage-defective mutant of flhB, the protein responsible for the switch to late substrate secretion, is completely abolished. FliK deletion variants that retain the ability to measure hook length are secreted thus demonstrating that FliK directly measures rod-hook length during the secretion process. Finally, we present a unifying model accounting for all published data on hook-length control in which FliK acts as a molecular ruler that takes measurements of rod-hook length while being intermittently secreted during the assembly process of the hook–basal body complex. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
4. Acetyl phosphate-sensitive regulation of flagellar biogenesis and capsular biosynthesis depends on the Rcs phosphorelay.
- Author
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Fredericks, Christine E., Shibata, Satoshi, Aizawa, Shin-Ichi, Reimann, Sylvia A., and Wolfe, Alan J.
- Subjects
ESCHERICHIA coli ,PHOSPHATES ,BIOSYNTHESIS ,CELLS ,GENETIC mutation - Abstract
As part of our attempt to map the impact of acetyl phosphate (acetyl∼P) on the entire network of two-component signal transduction pathways in Escherichia coli, we asked whether the influence of acetyl∼P on capsular biosynthesis and flagellar biogenesis depends on the Rcs phosphorelay. To do so, we performed a series of epistasis experiments: mutations in the components of the pathway that controls acetyl∼P levels were combined with mutations in components of the Rcs phosphorelay. Cells that did not synthesize acetyl∼P produced no capsule under normally permissive conditions, while those that accumulated acetyl∼P synthesized capsule under conditions previously considered to be non-permissive. Acetyl∼P-dependent capsular biosynthesis required both RcsB and RcsA, while the lack of RcsC restored capsular biosynthesis to acetyl∼P-deficient cells. Similarly, acetyl∼P-sensitive repression of flagellar biogenesis was suppressed by the loss of RcsB (but not of RcsA), while it was enhanced by the lack of RcsC. Taken together, these results show that both acetyl∼P-sensitive activation of capsular biosynthesis and acetyl∼P-sensitive repression of flagellar biogenesis require the Rcs phosphorelay. Moreover, they provide strong genetic support for the hypothesis that RcsC can function as either a kinase or a phosphatase dependent on environmental conditions. Finally, we learned that RcsB and RcsC inversely regulated the timing of flagellar biogenesis: rcsB mutants elaborated flagella prematurely, while rcsC mutants delayed their display of flagella. Temporal control of flagella biogenesis implicates the Rcs phosphorelay (and, by extension, acetyl∼P) in the transition of motile, planktonic individuals into sessile biofilm communities. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF
5. Mutations in Flk, FlgG, FlhA, and FlhE That Affect the Flagellar Type III Secretion Specificity Switch in Salmonella enterica.
- Author
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Hirano, Takanori, Mizuno, Shino, Aizawa, Shin-Ichi, and Hughes, Kelly T.
- Subjects
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MICROBIAL mutation , *GENETIC mutation , *BIOLOGICAL variation , *CHROMOSOME abnormalities , *FLAGELLARIACEAE , *EXCRETION , *BIOLOGICAL transport , *FOODBORNE diseases , *GENETICS ,SALMONELLA genetics - Abstract
Upon completion of the flagellar hook-basal body (HBB) structure, the flagellar type III secretion system switches from secreting rod/hook-type to filament-type substrates. The secretion specificity switch has been reported to occur prematurely (prior to HBB completion) in flk-null mutants (P. Aldridge, J. E. Karlinsey, E. Becker, F. F. Chevance, and K. T. Hughes, Mol. Microbiol. 60:630-643, 2006) and in distal rod gene gain-of-function mutants (flgG* mutants) that produce filamentous rod structures (F. F. Chevance, N. Takahashi, J. E. Karlinsey, J. Gnerer, T. Hirano, R. Samudrala, S. Aizawa, and K. T. Hughes, Genes Dev. 21:2326-2335, 2007). A fusion of β-lactamase (Bla) to the C terminus of the filament-type secretion substrate FlgM was used to select for mutants that would secrete FlgM-Bla into the periplasmic space and show ampicillin resistance (Apr). Apr resulted from null mutations in the flhE gene, C-terminal truncation mutations in the flhA gene, null and dominant mutations in the flk gene, and flgG* mutations. All mutant classes required the hook length control protein (FliK) and the rod cap protein (FlgJ) for the secretion specificity switch to occur. However, neither the hook (FlgE) nor the hook cap (FlgD) protein was required for premature FlgM-Bla secretion in the flgG* and flk mutant strains, but it was in the flhE mutants. Unexpectedly, when deletions of either flgE or flgD were introduced into flgG* mutant strains, filaments were able to grow directly on the filamentous rod structures. [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
6. Roles of Multiple Flagellins in Flagellar Formation and Flagellar Growth Post Bdelloplast Lysis in Bdellovibrio bacteriovorus
- Author
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Iida, Yoshiko, Hobley, Laura, Lambert, Carey, Fenton, Andrew K., Sockett, R. Elizabeth, and Aizawa, Shin-Ichi
- Subjects
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BDELLOVIBRIO bacteriovorus , *PROTEINS , *FLAGELLA (Microbiology) , *GENETIC mutation , *ELECTRON microscopy , *GENETIC polymorphisms - Abstract
Abstract: Bdellovibrio bacteriovorus cells have a single polar flagellum whose helical pitch and diameter characteristically change near the midpoint, resulting in a tapered wave. There are six flagellin genes in the genome: fliC1 to fliC6. Accordingly, the flagellar filament is composed of several similar flagellin species. We have used knockout mutants of each gene and analyzed the mutational effects on the filament length and on the composition and localization of each flagellin species in the filament by electron microscopy and one- and two-dimensional polyacrylamide gel electrophoresis. The location and amounts of flagellins in a filament were determined to be as follows: a small amount of FliC3 at the proximal end, followed by a large amount of FliC5, a large amount of FliC1, a small amount of FliC2 in this order, and a large amount of FliC6 at the distal end. FliC4 was present at a low level, but the location was not determined. Filament lengths of newly born progeny cells increased during prolonged incubation in nutrient-deficient buffer. The newly formed part of the elongated filament was composed of mainly FliC6. Reverse transcription PCR analysis of flagellar gene expression over 5 days in buffer showed that fliC gene expression tailed off over 5 days in the wild-type cells, but in the fliC5 mutant, expression of the fliC2, fliC4, and fliC6 genes was elevated on day 5, suggesting that they may be expressed to compensate for the absence of a major component, FliC5. [Copyright &y& Elsevier]
- Published
- 2009
- Full Text
- View/download PDF
7. Flagellar Formation in C-Ring-Defective Mutants by Overproduction of FliI, the ATPase Specific for Flagellar Type III Secretion.
- Author
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Konishi, Manabu, Kanbe, Masaomi, McMurry, Jonathan L., and Aizawa, Shin-Ichi
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CYTOPLASM , *BIOLOGICAL transport , *ADENOSINE triphosphatase , *GENOTYPE-environment interaction , *GENES , *GENETIC mutation , *BACTERIAL genetics , *BACTERIOLOGY - Abstract
The flagellar cytoplasmic ring (C ring), which consists of three proteins, FliG, FliM, and FliN, is located on the cytoplasmic side of the flagellum. The C ring is a multifunctional structure necessary for flagellar protein secretion, torque generation, and switching of the rotational direction of the motor. The deletion of any one of the fliG, fliM, and fliN genes results in a Fla - phenotype. Here, we show that the overproduction of the flagellum-specific ATPase FliI overcomes the inability of basal bodies with partial C-ring structures to produce complete flagella. Flagella made upon FliI overproduction were paralyzed, indicating that an intact C ring is essential for motor function. In FliN- or FliM-deficient mutants, flagellum production was about 10% of the wild-type level, while it was only a few percent in FliG-deficient mutants, suggesting that the size of partial C rings affects the extent of flagellation. For flagella made in C-ring mutants, the hook length varied considerably, with many being markedly shorter or longer than that of the wild type. The broad distribution of hook lengths suggests that defective C rings cannot control the hook length as tightly as the wild type even though FliK and FlhB are both intact. [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
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8. Autonomous and FliK-Dependent Length Control of the Flagellar Rod in Salmonella enterica.
- Author
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Takahashi, Noriko, Mizuno, Shino, Hirano, Takanori, Chevance, Fabienne F. V., Hughes, Kelly T., and Aizawa, Shin-Ichi
- Subjects
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SALMONELLA , *GENETIC mutation , *FOOD pathogens , *ENTEROBACTERIACEAE , *PATHOGENIC microorganisms , *MICROBIOLOGY - Abstract
Salmonella flgG point mutations produce filamentous rod structures whose lengths are determined by FliK. FliK length variants produce rods with lengths proportional to the corresponding FliK molecular size, suggesting that FliK controls the length of not only the hook but also the rod by the same molecular mechanism. [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
9. Temperature-Hypersensitive Sites of the Flagellar Switch Component FliG in Salmonella enterica Serovar Typhimurium.
- Author
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Mashimo, Takuji, Hashimoto, Manami, Yamaguchi, Shigeru, and Aizawa, Shin-Ichi
- Subjects
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MICROBIAL proteins , *FLAGELLA (Microbiology) , *SALMONELLA typhimurium , *GENETIC mutation , *EFFECT of temperature on microorganisms - Abstract
Three flagellar proteins, FliG, FliM, and FliN (FliGMN), are the components of the C ring of the flagellar motor. The genes encoding these proteins are multifunctional; they show three different phenotypes (Fla-, Mot-, and Che-), depending on the sites and types of mutations. Some of the Mot- mutants previously characterized are found to be motile. Reexamination of all Mot- mutants in fliGMN genes so far studied revealed that many of them are actually temperature sensitive (TS); that is, they are motile at 20°C but nonmotile at 37°C. There were two types of TS mutants: one caused a loss of function that was not reversed by a return to the permissive temperature (rigid TS), and the other caused a loss that was reversed (hyper-TS). The rigid TS mutants showed an all-or-none phenotype; that is, once a structure was formed, the structure and function were stable against temperature shifts. All of fliM and fliN and most of the fliG TS mutants belong to this group. On the other hand, the hyper-TS mutants (three of the fliG mutants) showed a temporal swimming/stop phenotype, responding to temporal temperature shifts when the structure was formed at a permissive temperature. Those hyper-TS mutation sites are localized in the C-terminal domain of the FliG molecules at sites that are different from the previously proposed functional sites. We discuss a role for this new region of FliG in the torque generation of the flagellar motor. [ABSTRACT FROM AUTHOR]
- Published
- 2007
- Full Text
- View/download PDF
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