Characterization of a human genomic DNA fragment which rescues defective lipid-linked oligosaccharide synthesis in a mutant G258 cell line isolated from the FM3A mouse mammary carcinoma cell line.
The G258 mutant cell line, isolated from the FM3A mouse mammary carcinoma cell line, is temperature-sensitive for both cell growth and asparagine-linked glycosylation due to mutation at a single location. The biochemical defect in the G258 mutant resides in the formation of lipid-linked oligosaccharide, presumably in one of the steps of GDP-mannose-dependent mannosylation (Y. Nishikawa, J. Cell. Physiol. 119, 260-266, 1984; Y. Nishikawa, Biochim. Biophys. Acta 1091, 135-140, 1991). In the present study, we transfected human genomic DNA fragments into the G258 mutant by the radiation hybrid method and isolated transformants (KK-1, -3 and -4) which showed recovery from both temperature-sensitive cell growth and asparagine-linked glycosylation. These transformants contained a common Alu-containing human DNA fragment (1.3 kb) which will be used as a marker for isolating the gene that complements the defect of lipid-liked oligosaccharide synthesis in the G258 mutant.
Dept. of Industrial Chemistry, School of Engineering, Tokai University, Kanagawa, Japan.
K
K
Kataoka
T
T
Takahashi
D
D
Ayusawa
Y
Y
Nishikawa
D020087:Alu Elements / Q000235:genetics; D000818:Animals; D001216:Asparagine / Q000378:metabolism; D001483:Base Sequence; D015139:Blotting, Southern; D002452:Cell Count; D002455:Cell Division; D002470:Cell Survival; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D018900:Dinucleotide Repeats / Q000235:genetics; D005816:Genetic Complementation Test; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D020084:Long Interspersed Nucleotide Elements / Q000235:genetics; D015674:Mammary Neoplasms, Animal / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D009154:Mutation; D009844:Oligosaccharides / Q000096:biosynthesis*; D017422:Sequence Analysis, DNA; D013696:Temperature; D014170:Transformation, Genetic; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism
D006023:Glycoproteins; D009844:Oligosaccharides; D001216:Asparagine; D004247:DNA