A conformational switch driven by phosphorylation regulates the activity of the evolutionarily conserved SNARE Ykt6.

Kaitlyn McGrath, Shivani Agarwal, Marco Tonelli, Mykola Dergai, Anthony L Gaeta, Andrew K Shum, Jessica Lacoste, Yongbo Zhang, Wenyu Wen, Daayun Chung, Grant Wiersum, Aishwarya Shevade, Sofia Zaichick, Damian B van Rossum, Ludmilla Shuvalova, Jeffrey N Savas, Sergei Kuchin, Mikko Taipale, Kim A Caldwell, Guy A Caldwell, Dirk Fasshauer, Gabriela Caraveo

Journal: Proceedings of the National Academy of Sciences of the United States of America 2021;118(12):e2016730118

PMID: 33723042

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Abstract

Ykt6 is a soluble -ethylmaleimide sensitive factor activating protein receptor (SNARE) critically involved in diverse vesicular fusion pathways. While most SNAREs rely on transmembrane domains for their activity, Ykt6 dynamically cycles between the cytosol and membrane-bound compartments where it is active. The mechanism that regulates these transitions and allows Ykt6 to achieve specificity toward vesicular pathways is unknown. Using a Parkinson's disease (PD) model, we found that Ykt6 is phosphorylated at an evolutionarily conserved site which is regulated by Ca signaling. Through a multidisciplinary approach, we show that phosphorylation triggers a conformational change that allows Ykt6 to switch from a closed cytosolic to an open membrane-bound form. In the phosphorylated open form, the spectrum of protein interactions changes, leading to defects in both the secretory and autophagy pathways, enhancing toxicity in PD models. Our studies reveal a mechanism by which Ykt6 conformation and activity are regulated with potential implications for PD.

Address: Department of Neurology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611.; The National Magnetic Resonance Facility at Madison, University of Wisconsin, Madison, WI 53706.; Department of Fundamental Neurosciences, University of Lausanne, 1005 Lausanne, Switzerland.; Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL 35401.; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada.; Integrated Molecular Structure Education and Research Center, Department of Chemistry, Northwestern University, Evanston, IL 60208.; Institutes of Biomedical Sciences, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China.; Center for Structural Genomics of Infectious Diseases, Northwestern University, Chicago, IL 60611.; Department of Biological Sciences, University of Wisconsin, Milwaukee, WI 53211.; Division of Experimental Pathology, Department of Pathology, Penn State College of Medicine, Hershey, PA 17033.; The Jake Gittlen Laboratories for Cancer Research, Penn State College of Medicine, Hershey, PA 17033.; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada.; Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada.; Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL 35401.; Department of Neurology, Center for Neurodegeneration and Experimental Therapeutics, Nathan Shock Center of Excellence in the Basic Biology of Aging, University of Alabama at Birmingham School of Medicine, Birmingham, AL 35249.; Department of Neurobiology, Center for Neurodegeneration and Experimental Therapeutics, Nathan Shock Center of Excellence in the Basic Biology of Aging, University of Alabama at Birmingham School of Medicine, Birmingham, AL 35249.; Department of Neurology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611; [email protected].

DOI: 10.1073/pnas.2016730118

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MeSH Terms: Amino Acids, Autophagy, Carrier Proteins, Cell Membrane, Conserved Sequence, Evolution, Molecular, Models, Molecular, Phosphorylation, Protein Binding, Protein Conformation, Protein Interaction Domains and Motifs, R-SNARE Proteins, Structure-Activity Relationship