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Journal of Biochemistry Advance Access published online on December 13, 2007
Journal of Biochemistry, doi:10.1093/jb/mvm232
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© 2007 The Japanese Biochemical Society

Expression of a novel 90-kDa protein, Lsd90, involved in the metabolism of very long-chain fatty acid-containing phospholipids in a mitosis-defective fission yeast mutant

Kazuaki Yokoyamaa,b,*, Miyuki Nakagawaa, Masayuki Satoha, Shigeaki Saitohc,+, Naoshi Dohmaed, Ayako Haradaa, Noriko Satoha, Ken Karasawaa, Koji Takiod,+, Mitsuhiro Yanagidab,c and Keizo Inouea

aFaculty of Pharmaceutical Sciences, Teikyo University, Suarashi, Sagamiko-machi, Sagamihara-shi, Kanagawa 229-0195, Japan.
bCREST, Japan Science and Technology Agency, Sanbancho Building, 3, Sanbancho Chiyoda-ku, Tokyo 102-0075, Japan.
cGraduate School of Biostudies, Kyoto University, Yoshida-Honmachi, Sakyo-ku, Kyoto 606-8501, Japan.
dBiomolecular Characterization Team, RIKEN, Wako-shi, Saitama 351-0198, Japan.

*To whom correspondence should be addressed: Dr. Kazuaki Yokoyama, Tel. 042-685-3731, Fax. 042-685-3731, e-mail: yokoyama{at}pharm.teikyo-u.ac.jp

Received August 8, 2007; Accepted November 15, 2007


  Abstract

The fission yeast lsd1/fas2 strain carries a temperature-sensitive mutation of the fatty-acid-synthase {alpha}-subunit, exhibiting an aberrant mitosis lsd phenotype, with accumulation of very-long-chain fatty-acid-containing phospholipid (VLCFA-PL). A novel 90-kDa protein, Lsd90 (SPBC16E9.16c), was found to be newly expressed in small particle-like structures in lsd1/fas2 cells under restrictive conditions. Two mismatches leading to a double frame shift were found between the sequence of the lsd90+ gene registered in the genomic database and the sequences determined experimentally at the amino acid, cDNA and genomic DNA levels. Unexpectedly, overexpression and disruption of the lsd90+ gene in either lsd1/fas2 or wild-type cells did not affect either cell growth or expression of the lsd phenotype. The amounts of VLCFA-PL that accumulated in lsd90-overexpressing lsd1/fas2 cells were significantly lower than those in lsd1/fas2 cells, suggesting the involvement of Lsd90 in the metabolism of VLCFA-PL.

Key Words: Fatty acid synthase mutant lsd1/fas2, Lsd90 (SPBC16E9.16c), proteome analysis, two-dimensional electrophoresis, very-long-chain fatty-acid-containing phospholipids

+Present address: Shigeaki Saitoh, Division of Cell Biology, Institute of Life Science, Kurume University, 1-1 Hyakunen-kohen, Kurume, Fukuoka 839-0864, Japan, Koji Takio, Biometal Science Laboratory, RIKEN SPring-8 Center, Harima Institute, 1-1-1, Kouto, Sayo, Hyogo 679-5148 Japan.


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