Differential localization to cytoplasm, nucleus or P-bodies of yeast PKA subunits under different growth conditions
Differential localization to cytoplasm, nucleus or P-bodies of yeast PKA subunits under different growth conditions
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DOI：
10.1016/j.ejcb.2009.08.005
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发表时间：
2010-04-01
影响因子：
6.6
通讯作者：
Portela, P.
中科院分区：
文献类型：
--
作者：
Tudisca, V.;Recouvreux, V.;Portela, P.
我们的目的是在这项工作中，进一步表征的复杂性和特异性的三种不同的异构体（Tpk 1，Tpk 2和Tpk 3）的催化和调节（Bcy 1）的PKA亚基在酿酒酵母。因此，分析了PKA亚基在活细胞中的亚细胞定位通过使用菌株携带UP（绿色荧光蛋白）融合到每个PKA亚基。在葡萄糖指数生长过程中，Bcy 1和Tpk 2均定位于细胞核，而Tpk 1和Tpk 3则表现出混合的核质定位模式。在甘油指数生长过程中和稳定期，PKA亚基主要表现为细胞质定位，其中Tpk 2和Tpk 3而不是Bcy 1与P-小体和EGP小体相关。Tpk 3在缺糖和高渗胁迫过程中积累到P-小体中Tpk 3的缺失改变了P-小体形成的动力学蛋白表达分析表明，每个Tpk的相对表达模式从发酵代谢的低水平变化到稳定期的高水平。Tpk水平的增加与Bcy 1水平产生不平衡我们的数据表明，酵母中通过PKA的信号传导特异性可能由Tpk的每种亚型的特定亚细胞定位介导（C）2009 Elsevier GmbH版权所有。
Our aim in this work was to further characterize the complexity and specificity of the three different isoforms (Tpk1, Tpk2 and Tpk3) of the catalytic and regulatory (Bcy1) subunits of PKA in Saccharomyces cerevisiae We thus analyzed the subcellular localization of the PKA subunits in living cells by using strains carrying UP (green fluorescent protein) fused to each PKA subunit. During exponential growth on glucose, both Bcy1 and Tpk2 localized in the nucleus, whereas Tpk1 and Tpk3 showed a mixed pattern of nucleo-cytoplasmic localization During exponential growth on glycerol and during stationary phase, the PKA subunits showed mostly cytoplasmic localization, with the peculiarity that Tpk2 and Tpk3 but not Bcy1, were found associated to P-bodies and EGP bodies. Tpk3 was accumulated Into P-bodies during glucose deprivation and hyper osmotic stress Deletion of Tpk3 altered the kinetics of P-body formation Analysis of protein expression showed that the relative expression pattern of each Tpk changes from low levels under fermentative metabolism to higher levels during stationary phase. The increase in Tpk levels produced an imbalance with Bcy1 levels Our data suggest that the signaling specificity through PKA in yeast could be mediated by a particular subcellular localization of each isoform of Tpk (C) 2009 Elsevier GmbH All rights reserved.