Chow VT et al. (2001) Biochem Genet "Phylogenetic relationships of the seven coat protein subunits of the coatomer complex, and comparative sequence analysis of murine xenin and proxenin."

Yeo WM, Lim DP, Sakharkar MK, Chow VT

[Biochem Genet, 2001]

The coatomer complex is involved in intracellular protein transport and comprises an assembly of seven polypeptide subunits designated alpha, beta, beta', gamma, delta, epsilon, and zeta COP. Rooted phylogenetic trees constructed from the full-length cDNA and amino acid sequences of 49 COP entities in different eukaryotes from yeast to man generally revealed striking conservation of each subunit through evolution. Both nucleotide and protein trees displayed close relationships between alpha and beta' subunits, between beta and gamma subunits, and between delta and zeta subunits, implying evolution from common ancestors as well as functional similarity. Interestingly, although 6 out of 7 epsilon-COP genes appeared to be grouped and related to the beta-COP genes, 4 out of 7 epsilon

Sayaka Miyazaki et al. (2010) East Asia Worm Meeting "Caenorhabditis elegans expressing human glycosphingolipids-A novel tool to elucidate pathology of O157 infection"

Kazuya Nomura, Daisuke Murata, Jun Fujii, Kazuhisa Iwabuchi, Shohei MItani, Eriko Kage Nakadai, Kazuko H Nomura, Hitoshi Nakayama, Sayaka Miyazaki

[East Asia Worm Meeting, 2010]

Verotoxin (VT)-producing Escherichia coli O157 induces hemolytic uremic syndrome. Several studies have demonstrated that VT shows its enterotoxicity via binding to the VT receptor globotriaosylceramide (Gb3/CD77). VT inhibits protein synthesis by catalytic inactivation of eukaryotic ribosomes and it is also recently noted that VT triggers apoptosis in many cell types. However, the apoptotic signaling network has not been well understood. Therefore, to study infection via VT, we are trying to generate a Caenorhabditis elegans (C. elegans) strain expressing human Gb3. In this study, we show that C. elegans transfected with human genes could be a useful tool to elucidate the pathogenic mechanisms of O157. The glycosphingolipids (GSLs) in C. elegans have a unique core structure, GlcNAcbeta1-3Manbeta1-4Glcbeta1-Cer found widely in invertebrates but not in vertebrates. The C. elegans genes, bre-3 and bre-5 encode a beta4-mannosyltransferase and beta3-N-acetylglucosaminyltransferase to control synthesis of the GSL core structure, respectively. C. elegans GSLs are built on Manbeta1-4Glcbeta1-Cer (ManCer) synthesized by bre-3, while vertebrate GSLs are built on Galbeta1-4Glcbeta1-Cer (LacCer) catalyzed by the beta4-galactosyltranseferases, beta4GalT-5 and -6. To generate a C. elegans strain expressing human Gb3, bre-5 deletion allele was isolated from trimethylpsoralen/ultraviolet mutagenized libraries. We used bre-5 mutants and bre-3 RNAi for transfecting the LacCer synthesizing enzyme to obtain worms producing LacCer but lacking GSLs synthesized on LacCer. To bre-3 RNAi-treated bre-5 mutants, cloned beta4GalT-5 cDNA linked with vha-6 promoter to drive intestine specific expression was injected, and the transgenic strains were isolated. By using a specific monoclonal antibody against LacCer (Iwabuchi K, FEBS Lett. 2009), we confirmed expression of LacCer accumulated in the worms. Transfection of the gene for Gb3 synthesis to this strain is underway in our laboratory.

VT1380

Caenorhabditis elegans

CB3298

Caenorhabditis elegans

hlb541

Mus musculus

PHENOTYPE: Male mutants show increased percent body fat when fed a standard chow diet. [provided by MGI curators]

hlb579

Mus musculus

PHENOTYPE: Male mutants exhibit increased percent body fat when fed a standard chow diet. [provided by MGI curators]

hlb583

Mus musculus

PHENOTYPE: Male mutants exhibit increased percent body fat when fed a standard chow diet. [provided by MGI curators]

hlb388

Mus musculus

PHENOTYPE: Mice with this mutation exhibit elevated fasting serum HDL cholesterol levels when fed a chow diet. [provided by MGI curators]

hlb497

Mus musculus

PHENOTYPE: Mice with this mutation exhibit elevated fasting plasma triglyceride levels when fed standard chow containing 6% fat. [provided by MGI curators]

Hlb476

Mus musculus

PHENOTYPE: Mice with this mutation exhibit elevated fasting plasma high-density lipoprotein (HDL) cholesterol when fed standard chow containing 6% fat. [provided by MGI curators]