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J. Biol. Chem., Vol. 281, Issue 12, 8110-8117, March 24, 2006

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The GTP-binding Protein YlqF Participates in the Late Step of 50 S Ribosomal Subunit Assembly in Bacillus subtilis^*

From the Department of Bioinformatics and Genomics, Graduate School of Information Science, Nara Institute of Science and Technology, 8916-5, Takayama, Ikoma, Nara 630-0101, Japan

Bacillus subtilis YlqF belongs to the Era/Obg subfamily of small GTP-binding proteins and is essential for bacterial growth. Here we report that YlqF participates in the late step of 50 S ribosomal subunit assembly. YlqF was co-fractionated with the 50 S subunit, depending on the presence of noncleavable GTP analog. Moreover, the GTPase activity of YlqF was stimulated specifically by the 50 S subunit in vitro. Dimethyl sulfate footprinting analysis disclosed that YlqF binds to a unique position in 23 S rRNA. Yeast two-hybrid data revealed interactions between YlqF and the B. subtilis L25 protein (Ctc). The interaction was confirmed by the pull-down assay of the purified proteins. Specifically, YlqF is positioned around the A-site and P-site on the 50 S subunit. Proteome analysis of the abnormal 50 S subunits that accumulated in YlqF-depleted cells showed that L16 and L27 proteins, located near the YlqF-binding domain, are missing. Our results collectively indicate that YlqF will organize the late step of 50 S ribosomal subunit assembly.

Received for publication, November 23, 2005 , and in revised form, January 23, 2006.

^* This work was supported by a grant-in-aid for Scientific Research on Priority Areas, Genome Biology, from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and Grant-in-aid for Scientific Research (A), 17201040, from the Japan Society for Promotion of Sciences. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Present address; Dept. of Microbiology, Graduate School of Medical Science, Kanazawa University, 13-1, Takara-Machi, Kanazawa, Ishikawa 920-8640, Japan.

² To whom correspondence should be addressed. Tel.: 81-743-72-5430; Fax: 81-743-72-5439; E-mail: nogasawa{at}bs.naist.jp.

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