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Cytoplasmic penetration and persistent infection of mammalian cells by polyglutamine aggregates

Nature Cell Biology volume 11, pages 219–225 (2009)Cite this article

Abstract

Sequence-specific nucleated protein aggregation is closely linked to the pathogenesis of most neurodegenerative diseases and constitutes the molecular basis of prion formation1. Here we report that fibrillar polyglutamine peptide aggregates can be internalized by mammalian cells in culture where they gain access to the cytosolic compartment and become co-sequestered in aggresomes together with components of the ubiquitin-proteasome system and cytoplasmic chaperones. Remarkably, these internalized fibrillar aggregates are able to selectively recruit soluble cytoplasmic proteins with which they share homologous but not heterologous amyloidogenic sequences, and to confer a heritable phenotype on cells expressing the homologous amyloidogenic protein from a chromosomal locus.

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Figure 1: Synthetic polyglutamine peptides form filamentous aggregates that are internalized by mammalian cells in culture.
Figure 2: Internalized polyglutamine peptide aggregates access the cytosolic compartment.
Figure 3: Internalized fibrillar aggregates induce homotypic but not heterotypic aggregation of cytoplasmic reporters.
Figure 4: Propagation of homotypic polyQ aggregation in cell culture.

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Acknowledgements

We thank J. Perrino and J. Mulholland for help with electron and confocal microscopy; K. Kirkegaard for comments on the manuscript and G. Chang and T. Hastie for statistical analysis. This research was supported in part by a grant from the NINDS and a Stanford Graduate Fellowship to P.-H.R.; R.M. is supported by the CNRS and the ANR contract no. ANR-06-BLAN-0266.

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Authors and Affiliations

  1. Department of Biology, Stanford University, Stanford, 94305-5020, CA, USA

    Pei-Hsien Ren, Jane E. Lauckner, Ioulia Kachirskaia & Ron R. Kopito

  2. Laboratoire d'Enzymologie et Biochimie Structurales, Centre National de la Recherche Scientifique, Gif-sur-Yvette, 91198, France

    Ronald Melki

  3. Department of Cell Biology and Physiology, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, 63110, Missouri, USA

    John E. Heuser

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  1. Pei-Hsien Ren
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Contributions

P.-H.R. contributed principally to the conception and design of the study, acquisition, analysis and interpretation of the data; J.E.L. and R.M. acquired, analysed and interpreted data and provided critical comments; I.K. contributed the experiment in Fig. 3d; J.E.H. contributed to the electron microscopic analysis and contributed substantially to the interpretation of the data; R.R.K. conceived and designed the study, and wrote and revised the manuscript.

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Correspondence to Ron R. Kopito.

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The authors declare no competing financial interests.

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Ren, PH., Lauckner, J., Kachirskaia, I. et al. Cytoplasmic penetration and persistent infection of mammalian cells by polyglutamine aggregates. Nat Cell Biol 11, 219–225 (2009). https://doi.org/10.1038/ncb1830

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