Swaminathan Swarna, Masek Tomas, Molin Claes, Pospisek Martin, Sunnerhagen Per
Department of Cell and Molecular Biology, Lundberg Laboratory, Göteborg University, S-405 30 Göteborg, Sweden.
Mol Biol Cell. 2006 Mar;17(3):1472-82. doi: 10.1091/mbc.e05-07-0632. Epub 2005 Dec 28.
Rck2 is a mitogen-activated protein kinase-activated protein kinase in yeast implicated in translational regulation. rck2Delta mutants are mildly sensitive to oxidative stress, a condition that causes dissociation of actively translating ribosomes (polysomes). In rck2Delta cells, polysomes are lost to an even higher degree than in the wild-type upon stress. Cells overexpressing the catalytically inactive rck2-kd allele are highly sensitive to oxidative stress. In such cells, dissociation of polysomes upon stress was instead greatly delayed. The protein synthesis rate decreased to a similar degree as in wild-type cells, however, indicating that in rck2-kd cells, the polysome complexes were inactive. Array analyses of total and polysome-associated mRNAs revealed major deregulation of the translational machinery in rck2 mutant cells. This involves transcripts for cytosolic ribosomal proteins and for processing and assembly of ribosomes. In rck2Delta cells, weakly transcribed mRNAs associate more avidly with polysomes than in wild-type cells, whereas the opposite holds true for rck2-kd cells. This is consistent with perturbed regulation of translation elongation, which is predicted to alter the ratio between mRNAs with and without strong entry sites at ribosomes. We infer that imbalances in the translational apparatus are a major reason for the inability of these cells to respond to stress.
Rck2是酵母中的一种丝裂原活化蛋白激酶激活的蛋白激酶,参与翻译调控。rck2Delta突变体对氧化应激轻度敏感,氧化应激会导致正在进行翻译的核糖体(多核糖体)解离。在rck2Delta细胞中,应激时多核糖体的丢失程度比野生型细胞更高。过表达催化失活的rck2-kd等位基因的细胞对氧化应激高度敏感。在这类细胞中,应激时多核糖体的解离反而大大延迟。然而,蛋白质合成速率下降到与野生型细胞相似的程度,这表明在rck2-kd细胞中,多核糖体复合物是无活性的。对总mRNA和与多核糖体相关的mRNA进行阵列分析,揭示了rck2突变体细胞中翻译机制的主要失调。这涉及胞质核糖体蛋白以及核糖体加工和组装的转录本。在rck2Delta细胞中,转录较弱的mRNA与多核糖体的结合比野生型细胞更紧密,而rck2-kd细胞则相反。这与翻译延伸调控受到干扰一致,预计这会改变核糖体上有无强进入位点的mRNA之间的比例。我们推断翻译装置的失衡是这些细胞无法应对应激的主要原因。