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Journal of Bacteriology, February 2002, p. 983-991, Vol. 184, No. 4
0021-9193/01/$04.00+0     DOI: 10.1128/jb.184.4.983-991.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of Two myo-Inositol Transporter Genes of Bacillus subtilis

Ken-Ichi Yoshida, Yoshiyuki Yamamoto, Kaoru Omae, Mami Yamamoto, and Yasutaro Fujita*

Department of Biotechnology, Fukuyama University, Fukuyama, Hiroshima 729-0292, Japan

Received 31 July 2001/ Accepted 19 November 2001

Among hundreds of mutants constructed systematically by the Japanese groups participating in the functional analysis of the Bacillus subtilis genome project, we found that a mutant with inactivation of iolT (ydjK) exhibited a growth defect on myo-inositol as the sole carbon source. The putative product of iolT exhibits significant similarity with many bacterial sugar transporters in the databases. In B. subtilis, the iolABCDEFGHIJ and iolRS operons are known to be involved in inositol utilization, and its transcription is regulated by the IolR repressor and induced by inositol. Among the iol genes, iolF was predicted to encode an inositol transporter. Inactivation of iolF alone did not cause such an obvious growth defect on inositol as the iolT inactivation, while simultaneous inactivation of the two genes led to a more severe defect than the single iolT inactivation. Determination of inositol uptake by the mutants revealed that iolT inactivation almost completely abolished uptake, but uptake by IolF itself was slightly detectable. These results, as well as the Km and Vmax values for the IolT and IolF inositol transporters, indicated that iolT and iolF encode major and minor inositol transporters, respectively. Northern and primer extension analyses of iolT transcription revealed that the gene is monocistronically transcribed from a promoter likely recognized by {varsigma}sgr;A RNA polymerase and negatively regulated by IolR as well. The interaction between IolR and the iolT promoter region was analyzed by means of gel retardation and DNase I footprinting experiments, it being suggested that the mode of interaction is quite similar to that found for the promoter regions of the iol divergon.

* Corresponding author. Mailing address: Department of Biotechnology, Faculty of Engineering, Fukuyama University, 985 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 729-0292, Japan. Phone: 81 849-36-2111. Fax: 81 849-36-2459. E-mail: yfujita{at}bt.fubt.fukuyama-u.ac.jp.

Journal of Bacteriology, February 2002, p. 983-991, Vol. 184, No. 4
0021-9193/01/$04.00+0     DOI: 10.1128/jb.184.4.983-991.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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