Dictyostelium gnt15 encodes a protein with similarity to LARGE and plays an essential role in development.
LARGE is a putative glycosyltransferase found to be mutated in mice with myodystrophy or patients with congenital muscular dystrophy. By homology searches, we identified in the Dictyostelium discoideum genome four open reading frames, i.e. gnt12-15, encoding proteins with sequence similarity to LARGE. Semi-quantitative RT-PCR analysis revealed distinct temporal expression patterns of the four gnt genes throughout Dictyostelium development. To explore the gene function, we performed targeted disruptions of gnt14 and gnt15. The gnt14(-) strains showed no obvious phenotypes. However, gnt15(-) cells grew slowly, changed in morphology, and displayed a developmental phenotype arresting at early stages. Compared with the wild type, gnt15(-) cells were more adhesive and exhibited altered levels of some surface adhesion molecules. Moreover, lectin-binding analysis demonstrated that gnt15 disruption affected profiles of membrane glycoproteins. Taken together, our data suggest that Gnt15 is essential for Dictyostelium development and may have a role in modulating cell adhesion and glycosylation.
Pang
Te-Ling
TL
Institute of Biochemistry and Molecular Biology, School of Life Sciences, National Yang-Ming University, Taipei 11221, Taiwan.
Wu
Chao-Jung
CJ
Chen
Pin-An
PA
Weng
Yi-Lan
YL
Chen
Mei-Yu
MY
D000818:Animals; D019521:Body Patterning / Q000235:genetics*; D004023:Dictyostelium / Q000235:genetics* / Q000254:growth & development*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D009154:Mutation; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*
C502747:Large1 protein, mouse; D017351:N-Acetylglucosaminyltransferases