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Journal of Biochemistry Advance Access originally published online on December 13, 2007
Journal of Biochemistry 2008 143(3):369-375; 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 Yokoyama1,2,{dagger}, Miyuki Nakagawa1, Masayuki Satoh1, Shigeaki Saitoh3,{ddagger}, Naoshi Dohmae4, Ayako Harada1, Noriko Satoh1, Ken Karasawa1, Koji Takio4,{ddagger}, Mitsuhiro Yanagida2,3 and Keizo Inoue1

1Faculty of Pharmaceutical Sciences, Teikyo University, Suarashi, Sagamiko-machi, Sagamihara-shi, Kanagawa 229-0195; 2CREST, Japan Science and Technology Agency, Sanbancho Building, 3, Sanbancho Chiyoda-ku, Tokyo 102-0075; 3Graduate School of Biostudies, Kyoto University, Yoshida-Honmachi, Sakyo-ku, Kyoto 606-8501; and 4Biomolecular Characterization Team, RIKEN, Wako-shi, Saitama 351-0198, Japan

{dagger}To whom correspondence should be addressed. 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 sequences 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

Abbreviations: EMM, Edinburgh minimal medium; 2ME, 2-mercaptoethanol; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PVDF, polyvinylidene difluoride; VLCFA-PLs, very-long-chain fatty-acid-containing phospholipids; YPD, yeast extract peptone dextrose


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