TY - JOUR
T1 - CdsA is involved in biosynthesis of glycolipid MPIase essential for membrane protein integration in vivo
AU - Sawasato, Katsuhiro
AU - Sato, Ryo
AU - Nishikawa, Hanako
AU - Iimura, Naoki
AU - Kamemoto, Yuki
AU - Fujikawa, Kohki
AU - Yamaguchi, Toshiyuki
AU - Kuruma, Yutetsu
AU - Tamura, Yasushi
AU - Endo, Toshiya
AU - Ueda, Takuya
AU - Shimamoto, Keiko
AU - Nishiyama, Ken ichi
N1 - Funding Information:
We thank Prof Toshihiko Oka (Rikkyo University) for the discussion, Prof Andreas Kuhn (University of Hohenheim) for the plasmids, Ms Mari Saikudo (Iwate University) for the technical help, and NBRP (NIG, Japan): E.coli and E. coli Genetic Resources at Yale (CGSC) for E. coli strains. The experiments involving radioisotopes were carried out at the Radioisotope Laboratory of Iwate University. This work was supported by JSPS KAKENHI (Grant Numbers 17K13262 to K.F.; 16H06156, 16H00797, 26119704 and 26106003 to Y.Kuruma; 15H05595 to Y.T., 15H05705 and 22227003 to T.E.; 16H02089 to T.U.; 16H01166 to K.Shimamoto; 25291009, 26102703, 26119701, 15KT0073, 16H01374, 16K15083 and 17H02209 to K.N.) and JST CREST (Grant Number JPMJCR12M1 to T.E.).
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - MPIase is a glycolipid that is involved in membrane protein integration. Despite evaluation of its functions in vitro, the lack of information on MPIase biosynthesis hampered verification of its involvement in vivo. In this study, we found that depletion of CdsA, a CDP-diacylglycerol synthase, caused not only a defect in phospholipid biosynthesis but also MPIase depletion with accumulation of the precursors of both membrane protein M13 coat protein and secretory protein OmpA. Yeast Tam41p, a mitochondrial CDP-diacylglycerol synthase, suppressed the defect in phospholipid biosynthesis, but restored neither MPIase biosynthesis, precursor processing, nor cell growth, indicating that MPIase is essential for membrane protein integration and therefore for cell growth. Consistently, we observed a severe defect in protein integration into MPIase-depleted membrane vesicles in vitro. Thus, the function of MPIase as a factor involved in protein integration was proven in vivo as well as in vitro. Moreover, Cds1p, a eukaryotic CdsA homologue, showed a potential for MPIase biosynthesis. From these results, we speculate the presence of a eukaryotic MPIase homologue.
AB - MPIase is a glycolipid that is involved in membrane protein integration. Despite evaluation of its functions in vitro, the lack of information on MPIase biosynthesis hampered verification of its involvement in vivo. In this study, we found that depletion of CdsA, a CDP-diacylglycerol synthase, caused not only a defect in phospholipid biosynthesis but also MPIase depletion with accumulation of the precursors of both membrane protein M13 coat protein and secretory protein OmpA. Yeast Tam41p, a mitochondrial CDP-diacylglycerol synthase, suppressed the defect in phospholipid biosynthesis, but restored neither MPIase biosynthesis, precursor processing, nor cell growth, indicating that MPIase is essential for membrane protein integration and therefore for cell growth. Consistently, we observed a severe defect in protein integration into MPIase-depleted membrane vesicles in vitro. Thus, the function of MPIase as a factor involved in protein integration was proven in vivo as well as in vitro. Moreover, Cds1p, a eukaryotic CdsA homologue, showed a potential for MPIase biosynthesis. From these results, we speculate the presence of a eukaryotic MPIase homologue.
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U2 - 10.1038/s41598-018-37809-8
DO - 10.1038/s41598-018-37809-8
M3 - Article
C2 - 30718729
AN - SCOPUS:85061056569
SN - 2045-2322
VL - 9
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 1372
ER -