Microarray analysis confirms the specificity of a Chlamydomonas reinhardtii chloroplast RNA stability mutant
Microarray analysis confirms the specificity of a Chlamydomonas reinhardtii chloroplast RNA stability mutant
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DOI：
10.1104/pp.104.053256
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发表时间：
2005-02-01
期刊：
影响因子：
7.4
通讯作者：
Higgs, DC
中科院分区：
文献类型：
--
作者：
Erickson, B;Stern, DB;Higgs, DC
叶绿体和线粒体基因的表达依赖于核编码的蛋白质，其中一些控制细胞器RNA的加工，稳定性和/或翻译。为了测试一个这样的RNA稳定因子的特异性，我们使用了两个已知的莱茵衣藻非光合突变体携带突变的Mcd 1核基因（mcd 1 -1和mcd 1 -2）。我们以前报道过，这些突变体不能积累叶绿体petD mRNA及其产物，细胞色素B（6）/f复合物的亚基IV，这是光合作用所必需的。这种突变体通常被认为是基因特异性的，但没有严格测试。在这里，我们已经使用微阵列分析，以评估叶绿体，线粒体和核RNA的变化，因为很少有其他RNA显着改变这些突变体，得出结论，Mcd 1确实是专门需要petD mRNA积累。此外，在mcd 1 -2中发现了一个新的非连锁核突变，该突变大大降低了叶绿体atpA mRNA的积累。遗传分析表明，mda 1-ncc 1不能补充，其中含atpA的转录物也受到类似的影响（D. Drapier，J. Girard-Bascou，D. B. Stern，F. A. Wollman [2002] Plant J 31：687-697），并且我们将该推定的新等位基因命名为mda 1 -2。我们的结论是，DNA微阵列是有效的和有用的表征细胞器RNA积累突变体的特异性。
The expression of chloroplast and mitochondrial genes depends on nucleus-encoded proteins, some of which control processing, stability, and/or translation of organellar RNAs. To test the specificity of one such RNA stability factor, we used two known Chlamydomonas reinhardtii nonphotosynthetic mutants carrying mutations in the Mcd1 nuclear gene (mcd1-1 and mcd1-2). We previously reported that these mutants fail to accumulate the chloroplast petD mRNA and its product, subunit IV of the cytochrome b(6)/f complex, which is essential for photosynthesis. Such mutants are generally presumed to be gene specific but are not tested rigorously. Here, we have used microarray analysis to assess changes in chloroplast, mitochondrial, and nuclear RNAs, and since few other RNAs were significantly altered in these mutants, conclude that Mcd1 is indeed specifically required for petD mRNA accumulation. In addition, a new unlinked nuclear mutation was discovered in mcd1-2, which greatly reduced chloroplast atpA mRNA accumulation. Genetic analyses showed failure to complement mda1-ncc1, where atpA-containing transcripts are similarly affected (D. Drapier, J. Girard-Bascou, D.B. Stern, F.-A. Wollman [2002] Plant J 31: 687-697), and we have named this putative new allele mda1-2. We conclude that DNA microarrays are efficient and useful for characterizing the specificity of organellar RNA accumulation mutants.