Bun-Ya M, Harashima S, Oshima Y
Department of Biotechnology, Faculty of Engineering, Osaka University, Japan.
Mol Cell Biol. 1992 Jul;12(7):2958-66. doi: 10.1128/mcb.12.7.2958-2966.1992.
We have found an open reading frame which is 1.1 kb upstream of PHO84 (which encodes a Pi transporter) and is transcribed from the opposite strand. In Saccharomyces cerevisiae, this gene is distal to the TUB3 locus on the left arm of chromosome XIII and is named GTR1. GTR1 encodes a protein consisting of 310 amino acid residues containing, in its N-terminal region, the characteristic tripartite consensus elements for binding GTP conserved in GTP-binding proteins, except for histidine in place of a widely conserved aspargine residue in element III. Disruption of the GTR1 gene resulted in slow growth at 30 degrees C and no growth at 15 degrees C; other phenotypes resembled those of pho84 mutants and included constitutive synthesis of repressible acid phosphatase, reduced Pi transport activity, and resistance to arsenate. The latter phenotypes were shown to be due to a defect in Pi uptake, and the Gtr1 protein was found to be functionally associated with the Pho84 Pi transporter. Recombination between chromosome V (at the URA3 locus) and chromosome XIII (in the GTR1-PHO84-TUB3 region) by using a plasmid-encoded site-specific recombination system indicated that the order of these genes was telomere-TUB3-PHO84-GTR1-CENXIII.
我们发现了一个开放阅读框,它位于PHO84(编码一种磷酸盐转运蛋白)上游1.1 kb处,且从相反链转录。在酿酒酵母中,该基因位于第十三条染色体左臂上的TUB3基因座远端,被命名为GTR1。GTR1编码一种由310个氨基酸残基组成的蛋白质,其N端区域含有GTP结合蛋白中保守的用于结合GTP的特征性三联体共有元件,但元件III中广泛保守的天冬酰胺残基被组氨酸取代。GTR1基因的破坏导致在30℃下生长缓慢,在15℃下无法生长;其他表型与pho84突变体相似,包括可阻遏酸性磷酸酶的组成型合成、磷酸盐转运活性降低以及对砷酸盐的抗性。后一种表型被证明是由于磷酸盐摄取缺陷所致,并且发现Gtr1蛋白在功能上与Pho84磷酸盐转运蛋白相关。通过使用质粒编码的位点特异性重组系统,在第五条染色体(URA3基因座处)和第十三条染色体(GTR1 - PHO84 - TUB3区域)之间进行重组,结果表明这些基因的顺序是端粒 - TUB3 - PHO84 - GTR1 - 第十三条染色体着丝粒。