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Biomedical subjects

Akiko Kusumoto

Publications and source records attributed to Akiko Kusumoto.

6 recordsLinked to original sources
PubMed2006

Regulation of polar flagellar number by the flhF and flhG genes in Vibrio alginolyticus.

The number and location of bacterial flagella vary with the species. The Vibrio alginolyticus cell has a single polar flagellum, which is driven by sodium ions. We selected mutants on the basis of reduced swarming ability on soft agar plates. Among them, we found two mutants with multiple polar flagella, and named them KK148 and NMB155. In Pseudomonas species, it is known that FlhF and FleN, which are FtsY and MinD homologs, respectively, are involved in regulation of flagellar placement and number, respectively. We cloned homologous genes of V. alginolyticus, flhF and flhG. KK148 cells had a nonsense mutation in flhG; cells expressing transgenic flhG recovered the swarming ability and had a reduced number of polar flagella. NMB155 cells did not have a mutation in either flhF or flhG. In wild-type cells, expression of flhF increased the number of polar flagella; in contrast, expression of flhG reduced both the number of polar flagella and the swarming ability. These results suggest that FlhG negatively regulates the number of polar flagella in V. alginolyticus. KK148 cells expressing both flhF and flhG exhibited fewer polar flagella and better swarming ability than KK148 cells expressing flhG alone, suggesting that FlhG acts with FlhF.

Amino Acid Sequence↗
PubMed2006

Improving syllable identification by a preprocessing method reducing overlap-masking in reverberant environments.

Overlap-masking degrades speech intelligibility in reverberation [R. H. Bolt and A. D. MacDonald, J. Acoust. Soc. Am. 21(6), 577-580 (1949)]. To reduce the effect of this degradation, steady-state suppression has been proposed as a preprocessing technique [Arai et al., Proc. Autumn Meet. Acoust. Soc. Jpn., 2001; Acoust. Sci. Tech. 23(8), 229-232 (2002)]. This technique automatically suppresses steady-state portions of speech that have more energy but are less crucial for speech perception. The present paper explores the effect of steady-state suppression on syllable identification preceded by /a/ under various reverberant conditions. In each of two perception experiments, stimuli were presented to 22 subjects with normal hearing. The stimuli consisted of mono-syllables in a carrier phrase with and without steady-state suppression and were presented under different reverberant conditions using artificial impulse responses. The results indicate that steady-state suppression statistically improves consonant identification for reverberation times of 0.7 to 1.2 s. Analysis of confusion matrices shows that identification of voiced consonants, stop and nasal consonants, and bilabial, alveolar, and velar consonants were especially improved by steady-state suppression. The steady-state suppression is demonstrated to be an effective preprocessing method for improving syllable identification by reducing the effect of overlap-masking under specific reverberant conditions.

Adolescent↗
PubMed2005

Assembly of motor proteins, PomA and PomB, in the Na+-driven stator of the flagellar motor.

PomA and PomB are transmembrane proteins that form the stator complex in the sodium-driven flagellar motor of Vibrio alginolyticus and are believed to surround the rotor part of the flagellar motor. We constructed and observed green fluorescent protein (GFP) fusions of the stator proteins PomA and PomB in living cells to clarify how stator proteins are assembled and installed into the flagellar motor. We were able to demonstrate that GFP-PomA and GFP-PomB localized to a cell pole dependent on the presence of the polar flagellum. Localization of the GFP-fused stator proteins required their partner subunit, PomA or PomB, and the C-terminal domain of PomB, which has a peptidoglycan-binding motif. Each of the GFP-fused stator proteins was co-isolated with its partner subunit from detergent-solubilized membrane. From these lines of evidence, we have demonstrated that the stator proteins are incorporated into the flagellar motor as a PomA/PomB complex and are fixed to the cell wall via the C-terminal domain of PomB.

Bacterial Proteins↗
PubMed2004

Cell surface differentiation of Mycoplasma mobile visualized by surface protein localization.

Mycoplasma mobile has a flask-shaped cell morphology and glides toward its tapered end at a rate of 3-7 cell lengths per s (2.0-4.5 microm s(-1)) by an unknown mechanism. Gliding requires that the surface of the cell is in contact with a solid substrate, such as glass or plastic. In order to characterize the nature of the outer surface of M. mobile, monoclonal antibodies were raised against intact cells and screened for their ability to recognize surface proteins. Four antibodies were identified and their protein targets were determined. One antibody recognized the Gli349 protein, which is known to be involved in glass binding and gliding. This antibody was also able to displace attached M. mobile cells from glass, suggesting that Gli349 is the major adhesion protein in M. mobile. The other three antibodies recognized members of the Mvsp family of proteins, which are presumably the major surface antigens of M. mobile. Immunofluorescence studies were performed to localize these proteins on the surface of M. mobile cells. Gli349 localized to the proximal region of the tapered part of the cell (the 'neck'), while the various Mvsp family members showed several distinct patterns of subcellular localization. MvspN and MvspO localized to the distal end of the tapered part of the cell (the 'head'), MvspK localized to the main part of the cell (the 'body'), and MvspI localized to both the head and body but not the neck. This analysis shows that M. mobile surprisingly expresses multiple versions of its major surface antigen at once but differentiates its surface by differential localization of the various paralogues.

Amino Acid Sequence↗
PubMed2004

Identification of a 349-kilodalton protein (Gli349) responsible for cytadherence and glass binding during gliding of Mycoplasma mobile.

Several mycoplasma species are known to glide in the direction of the membrane protrusion (head-like structure), but the mechanism underlying this movement is entirely unknown. To identify proteins involved in the gliding mechanism, protein fractions of Mycoplasma mobile were analyzed for 10 gliding mutants isolated previously. One large protein (Gli349) was observed to be missing in a mutant m13 deficient in hemadsorption and glass binding. The predicted amino acid sequence indicated a 348,758-Da protein that was truncated at amino acid residue 1257 in the mutant. Immunofluorescence microscopy with a monoclonal antibody showed that Gli349 is localized at the head-like protrusion's base, which we designated the cell neck, and immunoelectron microscopy established that the Gli349 molecules are distributed all around this neck. The number of Gli349 molecules on a cell was estimated by immunoblot analysis to be 450 +/- 200. The antibody inhibited both the hemadsorption and glass binding of M. mobile. When the antibody was used to treat gliding mycoplasmas, the gliding speed and the extent of glass binding were inhibited to similar extents depending on the concentration of the antibody. This suggested that the Gli349 molecule is involved not only in glass binding for gliding but also in movement. To explain the present results, a model for the mechanical cycle of gliding is discussed.

Bacterial Adhesion↗