| Autor: |
Zou, J., Scocca, J.R., Krag, S.S. |
| Zdroj: |
Archives of Biochemistry and Biophysics; March 1995, Vol. 317 Issue: 2 p487-496, 10p |
| Abstrakt: |
The gene gptencoding uridine diphosphate N-acetyl-D-glucosamine:dolichol phosphate N-acetylglucosaminylphosphoryltransferase (L-G1PT) was isolated by screening a Schizosaccharomyces pombegenomic DNA library in λ phage under low-stringency hybridization using the Saccharomyces cerevisiaegene ALG7 as probe. Sequencing 2.4 kb of S. pombeDNA 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. pombeprotein was 50% identical to the S. cerevisiaeprotein and 43% identical to the protein from Chinese hamster ovary (CHO) cells. Overexpression of the gptgene in S. pombecells 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 gptprobe. Northern analysis indicated a single 1.8-kb message is transcribed from the gptgene. The gptgene 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 gptORF or the CHO L-G1PT cDNA. The S. pombe gptgene was mapped to chromosome 2 near top1 and ade1. |
| Databáze: |
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