Cloning of the gene for monogalactosyldiacylglycerol synthase and its evolutionary origin.
Monogalactosyldiacylglycerol (MGDG) synthase (UDPgalactose:1,2-diacylglycerol 3-beta-D-galactosyltransferase; EC 2.4.1.46) catalyzes formation of MGDG, a major structural lipid of chloroplast. We cloned a cDNA for the synthase from cucumber cDNA library. The full-length cDNA clone was 2142 bp, and it contains a 1575-bp open reading frame encoding 525 aa. The open reading frame consists of the regions for a mature protein (422 aa; Mr of 46,552) and transit peptide to chloroplast (103 aa). Although the molecular weight of mature protein region matched that purified from cucumber cotyledons, it was quite different from those purified from spinach (approximately 20 kDa) reported by other groups. The mature region of the protein was expressed in Escherichia coli as a fusion protein with glutathione S-transferase. The expression in E. coli showed that the protein catalyzed MGDG synthesis very efficiently. Therefore, we concluded that the cDNA encodes MGDG synthase in cucumber. In addition, the deduced amino acid sequence of the MGDG synthase cDNA showed homology with MurG of Bacillus subtilis and E. coli, which encode a glycosyltransferase catalyzing the last step of peptidoglycan synthesis in bacteria. This sequence homology implies that the machinery of chloroplast membrane biosynthesis is evolutionarily derived from that of cell wall biosynthesis in bacteria. This is consistent with the endosymbiotic hypothesis of chloroplast formation.
Shimojima
M
M
Department of Biological Sciences, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama, Kanagawa, Japan.
Ohta
H
H
Iwamatsu
A
A
Masuda
T
T
Shioi
Y
Y
Takamiya
K
K
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D018553:Cucumis sativus / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005700:Galactosyltransferases / Q000235:genetics* / Q000378:metabolism; D008055:Lipids / Q000032:analysis; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D001425:Bacterial Outer Membrane Proteins; D008055:Lipids; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D005700:Galactosyltransferases; D017351:N-Acetylglucosaminyltransferases; C064057:UDP-N-acetylglucosamine-N-acetylmuramyl-(pentapeptide)pyrophosphoryl-undecaprenol N-acetylglucosamine transferase; C109990:1,2-diacylglycerol 3-beta-galactosyltransferase