Abstract
Rab GTPases are key regulators of membrane traffic. The Rab GTPase Ypt1 is essential for endoplasmic reticulum (ER)-Golgi traffic, intra-Golgi traffic, and the macroautophagy pathway. To identify effectors on the macroautophagy pathway, known autophagy-related genes (Atg genes) required for macroautophagy were tagged with GFP and screened for mislocalization in the ypt1-2 mutant. At the pre-autophagosomal structure (PAS), the localization of the serine/threonine kinase Atg1 was affected in the ypt1-2 mutant. We then used an in vitro binding assay to determine if Atg1 and Ypt1 physically interact with each other and co-immunoprecipitation experiments were performed to address if Atg1 preferentially interacts with the GTP-bound form of Ypt1.
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Acknowledgements
Salary support for J.W., S.C., S.D., and S.F.-N. was provided by the Howard Hughes Medical Institute.
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Wang, J., Cervantes, S., Davis, S., Ferro-Novick, S. (2015). Identifying a Rab Effector on the Macroautophagy Pathway. In: Li, G. (eds) Rab GTPases. Methods in Molecular Biology, vol 1298. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2569-8_10
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DOI: https://doi.org/10.1007/978-1-4939-2569-8_10
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2568-1
Online ISBN: 978-1-4939-2569-8
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