Identification of autophagosome-associated proteins and regulators by quantitative proteomic analysis and genetic screens

Jörn Dengjel, Maria Høyer-Hansen, Maria Nielsen, Tobias Eisenberg, Lea M Harder, Søren Schandorff, Thomas Farkas, Thomas Kirkegaard, Andrea C Becker, Sabrina Schroeder, Katja Vanselow, Emma Lundberg, Mogens M Nielsen, Anders R Kristensen, Vyacheslav Akimov, Jakob Bunkenborg, Frank Madeo, Marja Jäättelä, Jens S. Andersen

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Abstract

Autophagy is one of the major intracellular catabolic pathways, but little is known about the composition of autophagosomes. To study the associated proteins, we isolated autophagosomes from human breast cancer cells using two different biochemical methods and three stimulus types: amino acid deprivation or rapamycin or concanamycin A treatment. The autophagosome-associated proteins were dependent on stimulus, but a core set of proteins was stimulus-independent. Remarkably, proteasomal proteins were abundant among the stimulus-independent common autophagosome-associated proteins, and the activation of autophagy significantly decreased the cellular proteasome level and activity supporting interplay between the two degradation pathways. A screen of yeast strains defective in the orthologs of the human genes encoding for a common set of autophagosome-associated proteins revealed several regulators of autophagy, including subunits of the retromer complex. The combined spatiotemporal proteomic and genetic data sets presented here provide a basis for further characterization of autophagosome biogenesis and cargo selection.
Original languageEnglish
JournalMolecular and Cellular Proteomics
Volume11
Issue number3
Pages (from-to)M111.014035
ISSN1535-9476
DOIs
Publication statusPublished - 2012

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    Dengjel, J., Høyer-Hansen, M., Nielsen, M., Eisenberg, T., Harder, L. M., Schandorff, S., Farkas, T., Kirkegaard, T., Becker, A. C., Schroeder, S., Vanselow, K., Lundberg, E., Nielsen, M. M., Kristensen, A. R., Akimov, V., Bunkenborg, J., Madeo, F., Jäättelä, M., & Andersen, J. S. (2012). Identification of autophagosome-associated proteins and regulators by quantitative proteomic analysis and genetic screens. Molecular and Cellular Proteomics, 11(3), M111.014035. https://doi.org/10.1074/mcp.M111.014035