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Merging of multiple signals regulating Δ9 fatty acid desaturase gene transcription in Saccharomyces cerevisiae

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Abstract

Fatty acid desaturation, which requires molecular oxygen (O2) as an electron acceptor, is catalyzed by Δ9 fatty acid desaturase, which is encoded by OLE1 in Saccharomyces cerevisiae. Transcription of the OLE1 gene is repressed by unsaturated fatty acids (UFAs) and activated by hypoxia and low temperatures via the endoplasmic reticulum membrane protein Mga2p. We previously reported the isolation of the nfo3-1 (negative factor for O LE1) mutant, which exhibits enhanced expression of OLE1 in the presence of UFA and under aerobic conditions. In this work, we demonstrated that the NFO3 gene is identical to OLE1 and that the nfo3-1 mutation (renamed ole1-101) alters arginine-346, in the vicinity of the conserved histidine-rich motif essential for the catalytic function of the Ole1 protein, to lysine. The ratio of UFAs to total fatty acids in the ole1-101 mutant was 60%, compared to 75% in the wild type, suggesting that the reduction in relative levels of intracellular UFAs activates OLE1 transcription. However, in ole1-101 cells grown in the presence of oleic acid, the level of OLE1 expression remained high, although the relative amount of UFAs in the ole1-101 mutant cells was almost the same as that in wild-type cells growing under the same conditions. By contrast, when cells were grown with linoleic acid, which has a lower melting point than oleic acid, the elevation of the OLE1 expression level due to the ole1-101 mutation was almost completely suppressed. These observations suggest that the ole1-101 cells activate OLE1 transcription by sensing not only the intracellular UFA level, but also membrane fluidity or the nature of the UFA species itself. Furthermore, we found that not only the fatty acid-regulated (FAR) element but also the O 2 -regulated (O2R) element in the OLE1 promoter was involved in the activation of OLE1 transcription by the ole1-101 mutation, and that the effects of the low-oxygen signal and the ole1-101-generated signal on OLE1 expression were not additive. Taken together, these findings suggest that signals associated with hypoxia, low temperatures and intracellular UFA depletion activate OLE1 transcription by a common pathway.

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Acknowledgements

We thank T. Yoshida and M. Nakajima (International Center for Biotechnology, Osaka University) for GC analysis of fatty acids. This work was partially supported by a Grant-in-Aid for Scientific Research on Priority Areas (No. 12024213) to S. H. from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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  1. Y. Nakagawa

    Present address: Department of Applied Chemistry and Biotechnology, Yamanashi University, 4-3-11 Takeda, 400-8511, Kofu-shi, Yamanashi , Japan

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  1. Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, 565-0871, Suita-shi, Osaka, Japan

    Y. Nakagawa, A. Ueda, Y. Kaneko & S. Harashima

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  1. Y. Nakagawa
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  2. A. Ueda
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Correspondence to S. Harashima.

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Communicated by C. P. Hollenberg

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Nakagawa, Y., Ueda, A., Kaneko, Y. et al. Merging of multiple signals regulating Δ9 fatty acid desaturase gene transcription in Saccharomyces cerevisiae . Mol Gen Genomics 269, 370–380 (2003). https://doi.org/10.1007/s00438-003-0845-z

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