Structural basis of p62/SQSTM1 helical filaments, their presence in p62 bodies and role in cargo recognition in the cell

Jakobi, A. J.; Huber, S. T.; Mortensen, S. A.; Schultz, S. W.; Palara, A.; Kuhm, T.; Shrestha, B. K.; Larmark, T.; Hagen, W. J. H.; Wilmanns, M.; Johansen, T.; Brech, A.; Sachse, C.

doi:10.1101/671792

bioRxiv · 10.1101/671792

Structural basis of p62/SQSTM1 helical filaments, their presence in p62 bodies and role in cargo recognition in the cell

Jakobi, A. J.·Huber, S. T.·Mortensen, S. A.·Schultz, S. W.·Palara, A.·Kuhm, T.·Shrestha, B. K.·Larmark, T.·Hagen, W. J. H.·Wilmanns, M.·Johansen, T.·Brech, A.·Sachse, C.

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Abstract

p62/SQSTM1 is an autophagy receptor and signaling adaptor with an N-terminal PB1 domain that forms the scaffold of phase-separated p62 bodies in the cell. The molecular determinants that govern PB1 domain filament formation in vitro remain to be determined and the role of p62 filaments inside the cell is currently unclear. We determined four high-resolution cryo-EM structures of different human and Arabidopsis PB1 domain assemblies and observed a filamentous ultrastructure of phase-separated p62/SQSTM1 bodies using correlative cellular EM. We show that oligomerization or polymerization, driven by a double arginine finger in the PB1 domain, is a general requirement for lysosomal targeting of p62. Furthermore, the filamentous assembly state of p62 is required for autophagosomal processing of the p62-specific cargo KEAP1. Our results show that using such mechanisms, p62 filaments can be critical for cargo recognition and are an integral part of phase separated p62 bodies.

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Jakobi, A. J., Huber, S. T., Mortensen, S. A., Schultz, S. W., Palara, A., Kuhm, T., Shrestha, B. K., Larmark, T., Hagen, W. J. H., Wilmanns, M., Johansen, T., Brech, A., Sachse, C.. 2019-06-14. Structural basis of p62/SQSTM1 helical filaments, their presence in p62 bodies and role in cargo recognition in the cell. https://doi.org/10.1101/671792

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