Ždralević Maša, Guaragnella Nicoletta, Giannattasio Sergio
CNR, Istituto di Biomembrane e Bioenergetica, Via Amendola 165/A, 70126, Bari, Italy.
Methods Mol Biol. 2015;1265:321-31. doi: 10.1007/978-1-4939-2288-8_22.
Mitochondrial retrograde signaling is a mitochondria-to-nucleus communication pathway, conserved from yeast to humans, by which dysfunctional mitochondria relay signals that lead to cell stress adaptation in physiopathological conditions by changes in nuclear gene expression. The best comprehension of components and regulation of retrograde signaling have been obtained in Saccharomyces cerevisiae, where retrograde target gene expression is regulated by RTG genes. In this chapter, we describe the methods to measure mitochondrial retrograde pathway activation in yeast cells by monitoring the mRNA levels of RTG target genes, such as those encoding for peroxisomal citrate synthase, aconitase, and NAD(+)-specific isocitrate dehydrogenase subunit 1, as well as the phosphorylation status of Rtg1/3p transcriptional factor which controls RTG target gene transcription.
线粒体逆行信号传导是一条从酵母到人类都保守的线粒体到细胞核的通讯途径，功能失调的线粒体通过该途径传递信号，导致生理病理条件下细胞通过核基因表达的变化实现应激适应。在酿酒酵母中，对逆行信号传导的成分和调控有了最好的理解，其中逆行靶基因的表达由RTG基因调控。在本章中，我们描述了通过监测RTG靶基因的mRNA水平来测量酵母细胞中线粒体逆行途径激活的方法，这些靶基因如编码过氧化物酶体柠檬酸合酶、乌头酸酶和NAD(+)特异性异柠檬酸脱氢酶亚基1的基因，以及控制RTG靶基因转录的Rtg1/3p转录因子的磷酸化状态。
