Torelli Anna, Marieschi Matteo, Castagnoli Barbara, Zanni Corrado, Gorbi Gessica, Corradi Maria Grazia
Department of Evolutive and Functional Biology, University of Parma, Via Usberti 11/A, 43100 Parma, Italy.
Aquat Toxicol. 2008 Mar 26;86(4):495-507. doi: 10.1016/j.aquatox.2007.12.010. Epub 2008 Jan 8.
The gene expression of the wild type (S2-N) and a Cr-tolerant strain (S2-T) of the unicellular green alga Scenedesmus acutus has been compared in order to get more insight on their different chromium sensitivity. The RNA of the two strains was extracted after 4 days of culture in standard medium without chromium and analyzed by means of RNA differential display. The two strains showed differential gene transcription even in the absence of the heavy metal and six putatively differential amplicons were evidenced in the Cr-tolerant strain. Among the isolated amplicons, S2-T A63 was much more pronouncedly transcribed in the tolerant than in the wild type strain and was further characterized. S2-T A63 corresponding gene is present with the same copy number in the wild type and tolerant genomes and corresponds to an mRNA of about 2000 nt. The corresponding transcript is overexpressed in the Cr-tolerant strain after a 4-day culture and is not up-regulated by chromium exposure. The S2-T A63 sequence, obtained up to now, does not show significant homologies with any known gene. However, the analysis of the putative translation product reveals the presence of an interrupted fasciclin domain. This extracellular domain has been found in proteins from mammals, insects, echinoderms, plants, yeast and bacteria and is usually involved in cell adhesion. This finding suggests that the product for the S2-T A63 translation has an extracellular collocation, maybe as surface or secreted protein involved in external chromium detoxification.
为了更深入了解单细胞绿藻尖锐栅藻的野生型(S2-N)和耐铬菌株(S2-T)对铬敏感性的差异,对它们的基因表达进行了比较。在不含铬的标准培养基中培养4天后,提取了这两种菌株的RNA,并通过RNA差异显示进行分析。即使在没有重金属的情况下,这两种菌株也表现出不同的基因转录,并且在耐铬菌株中发现了六个推定的差异扩增子。在分离出的扩增子中,S2-T A63在耐铬菌株中的转录比野生型菌株中更为明显,并对其进行了进一步表征。S2-T A63对应的基因在野生型和耐铬基因组中的拷贝数相同,对应于一个约2000 nt的mRNA。经过4天的培养,相应的转录本在耐铬菌株中过表达,并且不受铬暴露的上调。到目前为止获得的S2-T A63序列与任何已知基因均无显著同源性。然而,对推定翻译产物的分析揭示了一个中断的成束蛋白结构域的存在。这种细胞外结构域已在来自哺乳动物、昆虫、棘皮动物、植物、酵母和细菌的蛋白质中发现,通常参与细胞粘附。这一发现表明,S2-T A63翻译产物具有细胞外定位,可能作为表面蛋白或分泌蛋白参与外部铬解毒。