Vaupotic Tomaz, Plemenitas Ana
Institute of Biochemistry, University of Ljubljana Faculty of Medicine, Vrazov Trg 2, SI-1000 Ljubljana, Slovenia.
BMC Genomics. 2007 Aug 16;8:280. doi: 10.1186/1471-2164-8-280.
Fluctuations in external salinity force eukaryotic cells to respond by changes in the gene expression of proteins acting in protective biochemical processes, thus counteracting the changing osmotic pressure. The high-osmolarity glycerol (HOG) signaling pathway is essential for the efficient up-regulation of the osmoresponsive genes. In this study, the differential gene expression of the extremely halotolerant black yeast Hortaea werneckii was explored. Furthermore, the interaction of mitogen-activated protein kinase HwHog1 and RNA polymerase II with the chromatin in cells adapted to an extremely hypersaline environment was analyzed.
A cDNA subtraction library was constructed for H. werneckii, adapted to moderate salinity or an extremely hypersaline environment of 4.5 M NaCl. An uncommon osmoresponsive set of 95 differentially expressed genes was identified. The majority of these had not previously been connected with the adaptation of salt-sensitive S. cerevisiae to hypersaline conditions. The transcriptional response in hypersaline-adapted and hypersaline-stressed cells showed that only a subset of the identified genes responded to acute salt-stress, whereas all were differentially expressed in adapted cells. Interaction with HwHog1 was shown for 36 of the 95 differentially expressed genes. The majority of the identified osmoresponsive and HwHog1-dependent genes in H. werneckii have not been previously reported as Hog1-dependent genes in the salt-sensitive S. cerevisiae. The study further demonstrated the co-occupancy of HwHog1 and RNA polymerase II on the chromatin of 17 up-regulated and 2 down-regulated genes in 4.5 M NaCl-adapted H. werneckii cells.
Extremely halotolerant H. werneckii represents a suitable and highly relevant organism to study cellular responses to environmental salinity. In comparison with the salt-sensitive S. cerevisiae, this yeast shows a different set of genes being expressed at high salt concentrations and interacting with HwHog1 MAP kinase, suggesting atypical processes deserving of further study.
外部盐度的波动迫使真核细胞通过参与保护性生化过程的蛋白质基因表达变化来做出反应,从而抵消不断变化的渗透压。高渗甘油(HOG)信号通路对于渗透压响应基因的有效上调至关重要。在本研究中,探索了极端耐盐黑酵母沃尔尼克霍塔酵母(Hortaea werneckii)的差异基因表达。此外,分析了丝裂原活化蛋白激酶HwHog1和RNA聚合酶II与适应极端高盐环境的细胞中染色质的相互作用。
构建了适应中等盐度或4.5M NaCl极端高盐环境的沃尔尼克霍塔酵母的cDNA消减文库。鉴定出一组95个差异表达的罕见渗透压响应基因。其中大多数以前未与盐敏感的酿酒酵母适应高盐条件相关联。高盐适应和高盐胁迫细胞中的转录反应表明,只有一部分鉴定出的基因对急性盐胁迫有反应,而所有基因在适应细胞中均有差异表达。95个差异表达基因中的36个显示与HwHog1相互作用。在沃尔尼克霍塔酵母中鉴定出的大多数渗透压响应和HwHog1依赖性基因以前在盐敏感的酿酒酵母中未被报道为Hog1依赖性基因。该研究进一步证明了在适应4.5M NaCl的沃尔尼克霍塔酵母细胞中,HwHog1和RNA聚合酶II在17个上调基因和2个下调基因的染色质上共同占据。
极端耐盐的沃尔尼克霍塔酵母是研究细胞对环境盐度反应的合适且高度相关的生物体。与盐敏感的酿酒酵母相比,这种酵母在高盐浓度下表达不同的基因集并与HwHog1丝裂原活化蛋白激酶相互作用,表明存在值得进一步研究的非典型过程。