Hildebrand M, Dahlin K, Volcani B E
Marine Biology Research Division and Center for Molecular Genetics, Scripps Institution of Oceanography, University of California San Diego, La Jolla 92093-0202, USA.
Mol Gen Genet. 1998 Dec;260(5):480-6. doi: 10.1007/s004380050920.
The transport of silicon is an integral part of the synthesis of the silicified cell wall of diatoms, yet knowledge of the number, features, and regulation of silicon transporters is lacking. We report the isolation and sequence determination of five silicon transporter (SIT) genes from Cylindrotheca fusiformis, and examine their expression patterns during cell wall synthesis. The encoded SIT amino acid sequences are highly conserved in their putative transmembrane domains. Nine conserved cysteines in this domain may account for the sensitivity of silicon uptake to sulfhydryl blocking agents. A less conserved C-terminal domain is predicted to form coiled-coil structures, suggesting that the SITs interact with other proteins. We show that SIT gene expression is induced just prior to, and during, cell wall synthesis. The genes are expressed at very different levels, and SIT1 is expressed in a different pattern from SIT 2-5. Hybridization experiments show that multiple SIT gene copies are present in all diatom species tested. From the data we infer that individual transporters play specific roles in silicon uptake, and propose that the cell regulates uptake by controlling the amount or location of each. The identification of all SIT genes in C. fusiformis will enhance our understanding of the mechanism and control of silicon transport in diatoms.
硅的转运是硅藻硅化细胞壁合成不可或缺的一部分,但目前对于硅转运蛋白的数量、特性及调控的了解仍很欠缺。我们报告了从梭形筒柱藻中分离并测定五个硅转运蛋白(SIT)基因的序列,并研究了它们在细胞壁合成过程中的表达模式。编码的SIT氨基酸序列在其假定的跨膜结构域中高度保守。该结构域中的九个保守半胱氨酸可能是硅摄取对巯基阻断剂敏感的原因。预测一个保守性较低的C末端结构域会形成卷曲螺旋结构,这表明SIT与其他蛋白质相互作用。我们发现SIT基因表达在细胞壁合成之前及过程中被诱导。这些基因的表达水平差异很大,且SIT1的表达模式与SIT2 - 5不同。杂交实验表明,在所有测试的硅藻物种中都存在多个SIT基因拷贝。从这些数据我们推断,单个转运蛋白在硅摄取中发挥特定作用,并提出细胞通过控制每个转运蛋白的数量或位置来调节摄取。确定梭形筒柱藻中的所有SIT基因将增进我们对硅藻中硅转运机制及调控的理解。