Asparagine-linked glycosylation in Schizosaccharomyces pombe: functional conservation of the first step in oligosaccharide-lipid assembly.
Asparagine-linked glycosylation in Schizosaccharomyces pombe: functional conservation of the first step in oligosaccharide-lipid assembly.
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粟酒裂殖酵母中天冬酰胺连接的糖基化：寡糖-脂质组装第一步的功能保守。
DOI：
10.1006/abbi.1995.1192
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Krag,SS
中科院分区：
文献类型：
--
作者：
Zou,J;Scocca,JR;Krag,SS
以酿酒酵母基因ALG 7为探针，通过低严格杂交筛选粟酒裂殖酵母λ噬菌体基因组DNA文库，获得了编码尿苷二磷酸N-乙酰-D-葡萄糖胺：多萜醇磷酸N-乙酰葡萄糖胺磷酸转移酶（L-G1 PT）的基因gpt。测序2.4kb的S. pombe DNA显示一个1338-bp的开放阅读框架（ORF），编码446个氨基酸的疏水蛋白，预测分子量为49，852。色葡萄pombe蛋白与S.酿酒酵母蛋白和43%相同的蛋白从中国仓鼠卵巢（CHO）细胞。gpt基因在S.粟酒裂殖酵母细胞对衣霉素的抗性增加了25倍，并且在分离的细胞膜中酶的比活性增加了13倍。这伴随着与gpt探针杂交的poly（A）+RNA增加50倍。北方分析表明，一个单一的1.8 kb的信息是从gpt基因转录。gpt基因对S.粟球。含有破坏的ORF的细胞可以通过含有完整的S.粟酒裂殖酵母gpt ORF或CHO L-G1 PT cDNA。色葡萄粟酒谷丙转氨酶基因定位于2号染色体上，靠近top1和ade 1。
The gene gpt encoding uridine diphosphate N-acetyl-D-glucosamine:dolichol phosphate N-acetylglucosaminylphosphoryltransferase (L-G1PT) was isolated by screening a Schizosaccharomyces pombe genomic DNA library in λ phage under low-stringency hybridization using the Saccharomyces cerevisiae gene ALG7 as probe. Sequencing 2.4 kb of S. pombe DNA revealed a 1338-bp open reading frame (ORF) encoding a hydrophobic protein of 446 amino acids with a predicted molecular weight of 49,852. The S. pombe protein was 50% identical to the S. cerevisiae protein and 43% identical to the protein from Chinese hamster ovary (CHO) cells. Overexpression of the gpt gene in S. pombe cells increased resistance to tunicamycin 25-fold and increased the specific activity of the enzyme in isolated cell membranes 13-fold. This was accompanied by a 50-fold increase in poly(A)+RNA hybridizing to the gpt probe. Northern analysis indicated a single 1.8-kb message is transcribed from the gpt gene. The gpt gene is essential for viability of S. pombe. Cells containing a disrupted ORF could be rescued by an expression plasmid containing either the intact S. pombe gpt ORF or the CHO L-G1PT cDNA. The S. pombe gpt gene was mapped to chromosome 2 near top1 and ade1.