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Journal of Bacteriology, May 2000, p. 2492-2497, Vol. 182, No. 9
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

An n-Alkane-Responsive Promoter Element Found in the Gene Encoding the Peroxisomal Protein of Candida tropicalis Does Not Contain a C6 Zinc Cluster DNA-Binding Motif

Tamotsu Kanai, Akihiro Hara, Naoki Kanayama, Mitsuyoshi Ueda, and Atsuo Tanaka*

Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan

Received 22 October 1999/Accepted 9 February 2000

When an asporogenic diploid yeast, Candida tropicalis, is cultivated on n-alkane, the expression of the genes encoding enzymes of the peroxisomal beta -oxidation pathway is highly induced. An upstream activation sequence (UAS) which can induce transcription in response to n-alkane (UASALK) was identified on the promoter region of the peroxisomal 3-ketoacyl coenzyme A (CoA) thiolase gene of C. tropicalis (CT-T3A). The 29-bp region (from -289 to -261) present upstream of the TATA sequence was sufficient to induce n-alkane-dependent expression of a reporter gene. Besides n-alkane, UASALK-dependent gene expression also occurred in the cells grown on oleic acid. Several kinds of mutant UASALK were constructed and tested for their UAS activity. It was clarified that the important nucleotides for UASALK activity were located within 10-bp region from -273 to -264 (5'-TCCTGCACAC-3'). This region did not contain a CGG triplet and therefore differed from the sequence of the oleate-response element (ORE), which is a UAS found on the promoter region of 3-ketoacyl-CoA thiolase gene of Saccharomyces cerevisiae. Similar sequences to UASALK were also found on several peroxisomal enzyme-encoding genes of C. tropicalis.

* Corresponding author. Mailing address: Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan. Phone: 81-75-753-5524. Fax: 81-75-753-5534. E-mail: atsuo{at}sbchem.kyoto-u.ac.jp.

Journal of Bacteriology, May 2000, p. 2492-2497, Vol. 182, No. 9
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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