Abstract
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.
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Acknowledgments
Rtg3p antibody was kindly provided by Dr. Z. Liu, University of New Orleans, New Orleans (USA). This work has been funded by grants from project FIRB-MERIT RBNE08HWLZ and the Italian Ministry of Economy and Finance to the CNR for the Project “FaReBio di Qualità.” M.Ž. is a recipient of a CNR Ph.D. fellowship in Biology and Biotechnologies, University of Salento, Lecce, Italy.
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Ždralević, M., Guaragnella, N., Giannattasio, S. (2015). Yeast as a Tool to Study Mitochondrial Retrograde Pathway En Route to Cell Stress Response. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine. Methods in Molecular Biology, vol 1265. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2288-8_22
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DOI: https://doi.org/10.1007/978-1-4939-2288-8_22
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