Ramon A M, Gil R, Burgal M, Sentandreu R, Valentin E
Sección de Microbiología Facultad de Farmacia, Universidad de Valencia, Spain.
Yeast. 1996 Dec;12(15):1535-48. doi: 10.1002/(SICI)1097-0061(199612)12:15%3C1535::AID-YEA59%3E3.0.CO;2-D.
A cDNA clone specifying a cell wall protein was isolated from a Yarrowia lipolytica cDNA library. The cDNA library was constructed in the expression vector lambda gt 11, with the RNA isolated from actively growing mycelial cells. The deduced amino acid sequence shows that the encoded protein contains an N-terminal hydrophobic signal peptide. We have designated this protein YWP1 for Yarrowia lipolytica cell Wall Protein. Northern hybridization identified YWP1 transcript only when Y. lipolytica was growing in the mycelial form. The encoded protein seems to be covalently bound to the glucan cell wall since it is not released from the cell walls by sodium dodecyl sulphate extraction, but it is solubilized following partial degradation of beta-glucan by Zymolyase digestion. The protein is localized in the outer surface on the tip of the growing mycelial cells and is found partially cryptic in sub-apical locations, suggesting that it participates directly in the mycelial wall architecture.
从解脂耶氏酵母cDNA文库中分离出一个编码细胞壁蛋白的cDNA克隆。该cDNA文库构建于表达载体λgt 11中,所用RNA取自活跃生长的菌丝体细胞。推导的氨基酸序列表明,编码的蛋白质含有一个N端疏水信号肽。我们将这种蛋白质命名为YWP1,即解脂耶氏酵母细胞壁蛋白。Northern杂交显示,只有当解脂耶氏酵母以菌丝体形式生长时,才能鉴定出YWP1转录本。编码的蛋白质似乎与葡聚糖细胞壁共价结合,因为它不能通过十二烷基硫酸钠提取从细胞壁中释放出来,但在经溶菌酶消化使β-葡聚糖部分降解后可溶解。该蛋白质定位于生长菌丝体细胞顶端的外表面,在亚顶端位置部分隐藏,这表明它直接参与菌丝体壁结构。