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J. Biol. Chem., Vol. 280, Issue 18, 18511-18516, May 6, 2005

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Leader Peptide-mediated Transcriptional Attenuation of Lysine Biosynthetic Gene Cluster in Thermus thermophilus^*

Taishi Tsubouchi, Reiko Mineki, Hikari Taka, Naoko Kaga, Kimie Murayama, Chiharu Nishiyama¶, Hisakazu Yamane, Tomohisa Kuzuyama, and Makoto Nishiyama||

From the Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan and BioMedical Research Center and ^¶Atopy Research Center, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan

The molecular mechanism for regulation of the genes involved in the biosynthesis of amino acids is poorly identified in Thermus thermophilus. In this study, we analyzed the transcriptional control of the major lysine biosynthetic gene cluster in T. thermophilus. S1 nuclease mapping revealed that the transcription, which is repressed by lysine, starts at 111 bp, upstream of the translational start codon, ATG, for the homocitrate synthase (hcs) gene. The 5'-leader region of 111 bp carries a sequence that can encode a short peptide of 14 amino acids with tandem-arranged lysine residues in its sequence. The nucleotide sequence of the region suggests that the transcript can form complicated secondary structures. Deletion of most of the 5'-leader region or mutation of the tandem lysine codons suppressed the transcriptional repression by lysine. Mutation of the tandem codons from lysine to glutamine resulted in glutamine-dependent repression of the gene connected downstream, indicating that the leader peptide mediated the transcriptional attenuation of the gene expression. This is the first report demonstrating the transcriptional regulation of amino acid biosynthesis in T. thermophilus.

Received for publication, December 22, 2004 , and in revised form, January 28, 2005.

^* This work was supported in part by a grant-in-aid for scientific research from the Ministry of Education, Culture, Sports, Science and Technology of Japan; the Noda Institute for Scientific Research; and the Nagase Science and Technology Foundation. 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.

^|| To whom correspondence should be addressed. Tel.: 81-3-5841-3074; Fax: 81-3-5841-8030; E-mail: umanis{at}mail.ecc.u-tokyo.ac.jp.

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				K. Fujiwara, T. Tsubouchi, T. Kuzuyama, and M. Nishiyama Involvement of the arginine repressor in lysine biosynthesis of Thermus thermophilus Microbiology, December 1, 2006; 152(12): 3585 - 3594. [Abstract] [Full Text] [PDF]