Guan Jiahn-Chou, Jinn Tsung-Luo, Yeh Ching-Hui, Feng Shi-Pin, Chen Yih-Ming, Lin Chu-Yung
Institute of Plant Biology, National Taiwan University, Taipei 106, Taiwan, ROC.
Plant Mol Biol. 2004 Nov;56(5):795-809. doi: 10.1007/s11103-004-5182-z. Epub 2005 Mar 24.
The cytosolic class I small heat shock proteins (sHSP-CI) represent the most abundant sHSP in plants. Here, we report the characterization and the expression profile of nine members of the sHSP-CI gene family in rice (Oryza sativa Tainung No.67), of which Oshsp16.9A, Oshsp16.9B, Oshsp16.9C, Oshsp16.9D and Oshsp17.9B are clustered on chromosome 1, and Oshsp17.3, Oshsp17.7, Oshsp17.9A and Oshsp18.0 are clustered on chromosome 3. Oshsp17.3 and Oshsp18.0 are linked by a 356-bp putative bi-directional promoter. Individual gene products were identified from the protein subunits of a heat shock complex (HSC) and from in vitro transcription/ translation products by two-dimensional gel electrophoreses (2-DE). All sHSP-CI genes except Oshsp17.9B were induced strongly after a 2-h heat shock treatment. The genes on chromosome 3 were induced rapidly at 32 and 41 degrees C, whereas those on chromosome 1 were induced slowly by similar conditions. Seven of these genes, except Oshsp16.9D and Oshsp17.9B, were induced by arsenite (As), but only genes on chromosome 3 were strongly induced by azetidine-2-carboxylic acid (Aze, a proline analog) and cadmium (Cd). A similar expression profile of all sHSP-CI genes at a lower level was evoked by ethanol, H2O2 and CuCl2 treatments. Transient expression assays of the promoter activity by linking to GUS reporter gene also supported the in vivo selective expression of the sHSP-CI genes by Aze treatment indicating the differential induction of rice sHSP-CI genes is most likely regulated at the transcriptional level. Only Oshsp16.9A abundantly accumulated in mature dry seed also suggested additionally prominent roles played by this HSP in development.
胞质I类小热激蛋白(sHSP-CI)是植物中最丰富的小热激蛋白。在此,我们报道了水稻(Oryza sativa Tainung No.67)中sHSP-CI基因家族9个成员的特征和表达谱,其中Oshsp16.9A、Oshsp16.9B、Oshsp16.9C、Oshsp16.9D和Oshsp17.9B聚集在1号染色体上,而Oshsp17.3、Oshsp17.7、Oshsp17.9A和Oshsp18.0聚集在3号染色体上。Oshsp17.3和Oshsp18.0由一个356 bp的假定双向启动子相连。通过二维凝胶电泳(2-DE)从热激复合体(HSC)的蛋白质亚基以及体外转录/翻译产物中鉴定出了各个基因产物。除Oshsp17.9B外,所有sHSP-CI基因在2小时热激处理后均被强烈诱导。3号染色体上的基因在32℃和41℃时迅速被诱导,而1号染色体上的基因在类似条件下诱导较慢。这些基因中的7个,除Oshsp16.9D和Oshsp17.9B外,可被亚砷酸盐(As)诱导,但只有3号染色体上的基因被氮杂环丁烷-2-羧酸(Aze,一种脯氨酸类似物)和镉(Cd)强烈诱导。乙醇、H2O2和CuCl2处理在较低水平上也引起了所有sHSP-CI基因相似的表达谱。通过与GUS报告基因连接进行启动子活性的瞬时表达分析也支持了Aze处理对sHSP-CI基因的体内选择性表达,表明水稻sHSP-CI基因的差异诱导很可能在转录水平受到调控。只有Oshsp16.9A在成熟干种子中大量积累,这也额外表明了该热激蛋白在发育过程中发挥的突出作用。