Molecular characterization of rice hsp101:: complementation of yeast hsp104 mutation by disaggregation of protein granules and differential expression in indica and japonica rice types
Molecular characterization of rice hsp101:: complementation of yeast hsp104 mutation by disaggregation of protein granules and differential expression in indica and japonica rice types
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DOI：
10.1023/a:1022324920316
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发表时间：
2003-03-01
影响因子：
5.1
通讯作者：
Grover, A
中科院分区：
文献类型：
--
作者：
Agarwal, M;Sahi, C;Grover, A
HSP 100蛋白是多种生物体热休克反应的重要组成部分。根据水稻hsp101基因cDNA序列设计特异性引物，对水稻hsp101基因进行PCR扩增和序列测定。Southern杂交分析表明，在水稻中每个单倍体基因组中似乎只有一个编码HSP 101蛋白的基因。北方分析表明热休克蛋白101转录本的表达是严格的热诱导和诱导是短暂的性质。在所测试的温度条件下，45 ℃处理2 h的完整水稻幼苗显示出最高水平的hsp101 mRNA。水稻全长hsp101 cDNA与插入突变体hsp104基因缺失的酵母突变体互补，其效果与拟南芥hsp101 cDNA相当。电子显微镜证据表明，水稻hsp101 cDNA在酵母中是积极的再溶解的胁迫诱导的蛋白颗粒在胁迫后的恢复期。水稻hsp101 cDNA在hsp104缺陷酵母中的表达也引起对砷的耐受性恢复。Western分析表明，这种蛋白质表达更迅速，在胁迫期间，并保留了更长的时间在胁迫后恢复期粳稻相比，籼稻类型。这是第一个报告，其中植物HSP 100蛋白表达与消失的蛋白颗粒在酵母细胞和不同的水稻类型依赖的蛋白表达模式的报告。
HSP100 protein is an important component of the heat-shock response in diverse organisms. Using specific primers based on cDNA sequence, rice hsp101 gene was PCR-amplified and sequenced. Southern analysis revealed that there appears to be a single gene per haploid genome coding for HSP101 protein in rice. Northern analysis showed that expression of hsp101 transcript is strictly heat-inducible and induction is transient in nature. In the temperature regime tested, 45 degreesC treatment to intact rice seedlings for 2 h showed maximal levels of hsp101 mRNA. Rice full-length hsp101 cDNA complemented yeast mutant disrupted for its own hsp104 gene by insertional mutagenesis, with efficacy that was comparable with Arabidopsis hsp101 cDNA. Electron micrographic evidence suggested that rice hsp101 cDNA in yeast is active in re-solubilizing the stress-induced protein granules in the post-stress recovery period. Rice hsp101 cDNA expression in hsp104 deficient yeast also caused recovery in tolerance against arsenite. Western analyses showed that this protein is expressed more rapidly during the stress period and retained for longer duration in the post-stress recovery period in japonica rice as compared to indica rice types. This is the first report wherein plant HSP100 protein expression is correlated to disappearance of protein granules in the yeast cells and distinct rice type-dependent protein expression patterns are reported.