Kearns M A, Monks D E, Fang M, Rivas M P, Courtney P D, Chen J, Prestwich G D, Theibert A B, Dewey R E, Bankaitis V A
Department of Cell Biology, University of Alabama at Birmingham, Birmingham, AL 35294-0005, USA.
EMBO J. 1998 Jul 15;17(14):4004-17. doi: 10.1093/emboj/17.14.4004.
Phosphatidylinositol transfer proteins (PITPs) have been shown to play important roles in regulating a number of signal transduction pathways that couple to vesicle trafficking reactions, phosphoinositide-driven receptor-mediated signaling cascades, and development. While yeast and metazoan PITPs have been analyzed in some detail, plant PITPs remain entirely uncharacterized. We report the identification and characterization of two soybean proteins, Ssh1p and Ssh2p, whose structural genes were recovered on the basis of their abilities to rescue the viability of PITP-deficient Saccharomyces cerevisiae strains. We demonstrate that, while both Ssh1p and Ssh2p share approximately 25% primary sequence identity with yeast PITP, these proteins exhibit biochemical properties that diverge from those of the known PITPs. Ssh1p and Ssh2p represent high-affinity phosphoinositide binding proteins that are distinguished from each other both on the basis of their phospholipid binding specificities and by their substantially non-overlapping patterns of expression in the soybean plant. Finally, we show that Ssh1p is phosphorylated in response to various environmental stress conditions, including hyperosmotic stress. We suggest that Ssh1p may function as one component of a stress response pathway that serves to protect the adult plant from osmotic insult.
磷脂酰肌醇转移蛋白(PITPs)已被证明在调节多种信号转导途径中发挥重要作用,这些信号转导途径与囊泡运输反应、磷酸肌醇驱动的受体介导的信号级联反应以及发育相关。虽然酵母和后生动物的PITPs已得到一定程度的详细分析,但植物PITPs仍完全未被表征。我们报告了两种大豆蛋白Ssh1p和Ssh2p的鉴定和表征,其结构基因是根据它们拯救PITP缺陷型酿酒酵母菌株活力的能力而获得的。我们证明,虽然Ssh1p和Ssh2p与酵母PITP的一级序列同源性约为25%,但这些蛋白表现出与已知PITPs不同的生化特性。Ssh1p和Ssh2p代表高亲和力的磷酸肌醇结合蛋白,它们在磷脂结合特异性以及在大豆植株中基本不重叠的表达模式方面相互区别。最后,我们表明Ssh1p在响应各种环境胁迫条件(包括高渗胁迫)时会发生磷酸化。我们认为Ssh1p可能作为应激反应途径的一个组成部分发挥作用,该途径有助于保护成年植株免受渗透损伤。