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. 1999 Aug;181(16):5103-6.
doi: 10.1128/JB.181.16.5103-5106.1999.

Functional interaction between PomA and PomB, the Na(+)-driven flagellar motor components of Vibrio alginolyticus

T Yorimitsu  1 , K Sato, Y Asai, I Kawagishi, M Homma
Affiliations

Functional interaction between PomA and PomB, the Na(+)-driven flagellar motor components of Vibrio alginolyticus

T Yorimitsu et al. J Bacteriol. 1999 Aug.

Abstract

Four proteins, PomA, PomB, MotX, and MotY, appear to be involved in force generation of the sodium-driven polar flagella of Vibrio alginolyticus. Among these, PomA and PomB seem to be associated and to form a sodium channel. By using antipeptide antibodies against PomA or PomB, we carried out immunoprecipitation to verify whether these proteins form a complex and examined the in vivo stabilities of PomA and PomB. As a result, we could demonstrate that PomA and PomB functionally interact with each other.

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Figures

FIG. 1
FIG. 1
Detection of proteins by immunoblotting with antipeptide antibodies against PomA (A) or PomB (B). Lanes 1, pSU41; lanes 2, pYA301; lanes 3, pYA303; lanes 4, pSK603. Molecular mass markers are indicated.
FIG. 2
FIG. 2
Immunoprecipitation assays with PomA1312 (A) and PomB93 (B). Lanes 1, NMB191 transformed with pSU41; lanes 2, VIO5; lanes 3, NMB155; lanes 4, NMB191 transformed with pYA303.
FIG. 3
FIG. 3
Stabilities of PomA and PomB. Overnight cultures of NMB191 cells with the pomA and/or pomB gene on a plasmid were inoculated at 1:50, and cells were harvested after 3, 6, 12, and 24 h. PomA or PomB was detected by immunoblotting with antibody PomA91 (A) or antibody PomB93 (B), respectively, as described in the text. Numbers below the lanes correspond to the times when cells were harvested.
FIG. 4
FIG. 4
Pulse-chase analysis of PomB. NMB191 cells harboring plasmids carrying pomA plus pomB (pYA303) (A) or pomB alone (pSK603) (B) were cultured in synthetic medium. At mid-log phase, Tran35S-label was added to 100 μCi/ml, and the mixture was incubated at 30°C for 30 min (pulse). Excess unlabeled methionine and cysteine were added to stop the labeling (chase at time zero). At the times indicated above the lanes, the cells were harvested by centrifugation, suspended in 1 ml of TNET buffer, and then immunoprecipitated with antibody PomB93, followed by SDS-PAGE and fluorography, as described in the text.
FIG. 5
FIG. 5
Coprecipitation of PomA and PomB. NMB191 cells transformed with the plasmid were labeled with Tran35S-label for 30 min and lysed, and the samples were immunoprecipitated either with antibody PomA1312 (A) or with antibody PomB93 (B) as described in the text. Samples were analyzed by SDS-PAGE followed by fluorography. The transformed plasmids were as follows: lanes 1, pSU41; lanes 2, pYA301; lanes 3, pSK603; lanes 4, pYA303.
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References

    1. Aizawa S-I. Flagellar assembly in Salmonella typhimurium. Mol Microbiol. 1996;19:1–5. - PubMed
    1. Asai, Y., I. Kawagishi, E. Sockett, and M. Homma. Hybrid motor with the H+- and Na+-driven components can rotate Vibrio polar flagella using sodium ions. Submitted for publication. - PMC - PubMed
    1. Asai Y, Kojima S, Kato H, Nishioka N, Kawagishi I, Homma M. Putative channel components for the fast-rotating sodium-driven flagellar motor of a marine bacterium. J Bacteriol. 1997;179:5104–5110. - PMC - PubMed
    1. Atsumi T, McCarter L, Imae Y. Polar and lateral flagellar motors of marine Vibrio are driven by different ion-motive forces. Nature (London) 1992;355:182–184. - PubMed
    1. Bartolomé B, Jubete Y, Martínez E, de la Cruz F. Construction and properties of a family of pACYC184-derived cloning vectors compatible with pBR322 and its derivatives. Gene. 1991;102:75–78. - PubMed

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