Yompakdee C, Bun-ya M, Shikata K, Ogawa N, Harashima S, Oshima Y
Department of Biotechnology, Faculty of Engineering, Osaka University, Japan.
Gene. 1996 May 24;171(1):41-7. doi: 10.1016/0378-1119(96)00079-0.
The PHO84 gene in Saccharomyces cerevisiae encodes the P(i) transporter Pho84p. The other three genes, GTR1, PHO86 and PHO87, are also suggested to be involved in the P(i) uptake system. We cloned and sequenced PHO86 and found that it encodes a 34-kDa protein consisting of 311 amino acid residues with two strongly hydrophobic segments in its N-terminal half. Western blotting analysis of cell extracts revealed that Pho86p, tagged with c-Myc, was fractionated into a water-insoluble fraction. Disruption of PHO86 did not affect cell viability even in combination with the pho84 and/or pho87 disruptions. The triple disruptants showed high levels of constitutive rAPase synthesis and arsenate resistance similar to the pho84 mutant, but showed slower cell growth than the pho84 mutant. PHO86 has two putative binding sites for the transcriptional activator, Pho4p, at nucleotide positions -191 and -497 relative to the ATG start codon, and showed substantial levels of transcription under high-P(i) conditions and more enhanced levels in low-P(i) medium.
酿酒酵母中的PHO84基因编码磷转运蛋白Pho84p。另外三个基因GTR1、PHO86和PHO87也被认为参与磷摄取系统。我们克隆并测序了PHO86,发现它编码一个由311个氨基酸残基组成的34 kDa蛋白,其N端一半有两个强疏水片段。对细胞提取物的蛋白质印迹分析表明,用c-Myc标记的Pho86p被分离到水不溶性部分。即使与pho84和/或pho87缺失组合,PHO86的缺失也不影响细胞活力。三重缺失突变体显示出与pho84突变体相似的高水平组成型rAPase合成和抗砷酸盐能力,但细胞生长比pho84突变体慢。PHO86在相对于ATG起始密码子的核苷酸位置-191和-497处有两个假定的转录激活因子Pho4p结合位点,在高磷条件下显示出相当水平的转录,在低磷培养基中水平更高。