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Titolo:
Cloning of the human cDNA which can complement the defect of the yeast mannosyltransferase I-deficient mutant alg 1
Autore:
Takahashi, T; Honda, R; Nishikawa, Y;
Indirizzi:
Tokai Univ, Sch Engn, Dept Ind Chem, Lab Glycobiol & Glycotechnol, Kanagawa 2591292, Japan Tokai Univ Kanagawa Japan 2591292 Glycotechnol, Kanagawa 2591292, Japan
Titolo Testata:
GLYCOBIOLOGY
fascicolo: 3, volume: 10, anno: 2000,
pagine: 321 - 327
SICI:
0959-6658(200003)10:3<321:COTHCW>2.0.ZU;2-U
Fonte:
ISI
Lingua:
ENG
Soggetto:
ASPARAGINE-LINKED GLYCOSYLATION; PHOSPHATE N-ACETYLGLUCOSAMINE-1-PHOSPHATE TRANSFERASE; PHOSPHOMANNOSE ISOMERASE DEFICIENCY; HAMSTER OVARY CELLS; SACCHAROMYCES-CEREVISIAE; ENDOPLASMIC-RETICULUM; PROTEIN GLYCOSYLATION; N-GLYCOSYLATION; ESCHERICHIA-COLI; OLIGOSACCHARIDE;
Keywords:
cDNA cloning; EST database; glycosylation; lipid-linked oligosaccharide; mannosyltransferase;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
47
Recensione:
Indirizzi per estratti:
Indirizzo: Takahashi, T Tokai Univ, Sch Engn, Dept Ind Chem, Lab Glycobiol & Glycotechnol, 1117 Kitakaname, Kanagawa 2591292, Japan Tokai Univ 1117 Kitakaname Kanagawa Japan 2591292 292, Japan
Citazione:
T. Takahashi et al., "Cloning of the human cDNA which can complement the defect of the yeast mannosyltransferase I-deficient mutant alg 1", GLYCOBIOLOG, 10(3), 2000, pp. 321-327

Abstract

The assembly of the lipid-linked oligosaccharide, Glc(3)Man(9)GlcNAc(2)-P-P-Dol, occurs on the rough ER membrane in an ordered stepwise manner, The process is highly conserved among eukaryotes, In order to isolate the human mannosyltransferase I (MT-I) gene involved in the process, we used the Saccharomyces cerevisiae MT-I gene (ALG1), which has already been cloned. On searching the EST database with the amino acid sequence of the ALG1 gene product, we detected seven related human EST clones. A human fetal brain cDNA library was screened by PCR using gene-specific primers based on the EST nucleotide sequences and a 430 bp cDNA fragment was amplified, The cDNA library was rescreened with this 430 bp cDNA, and two cDNA clones (HR1-3 and HR1-4) were isolated and sequenced. On a homology search of the EST database with the nucleotide sequence of HR1-3, we detected a novel human EST clone, AA675921 (GenBank accession number). Based on the nucleotide sequences of AA675921 and HR1-4, we designed gene-specific PCR primers, which allowed to amplify a 1.8 kb cDNA from human fetal brain cDNA, This cDNA was cloned and shown to contain an ORF encoding a protein of 464 amino acids. We designated this ORF as Hmat-1, The amino acid sequence deduced from the Hmat-1 gene showed several highly conserved regions shared with the yeast and nematode MT-I sequences. Furthermore, this 1.8 kb cDNA successfully complemented theS. cerevisiae alg1-1 mutation, indicating that the Hmat-1 gene encodes thehuman MT-P and that the function of this enzyme was conserved between yeast and human.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/12/20 alle ore 09:42:43