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Polyglutamine disruption of the huntingtin exon 1 N terminus triggers a complex aggregation mechanism

Nature Structural & Molecular Biology volume 16, pages 380–389 (2009)Cite this article

Abstract

Simple polyglutamine (polyQ) peptides aggregate in vitro via a nucleated growth pathway directly yielding amyloid-like aggregates. We show here that the 17-amino-acid flanking sequence (HTTNT) N-terminal to the polyQ in the toxic huntingtin exon 1 fragment imparts onto this peptide a complex alternative aggregation mechanism. In isolation, the HTTNT peptide is a compact coil that resists aggregation. When polyQ is fused to this sequence, it induces in HTTNT, in a repeat-length dependent fashion, a more extended conformation that greatly enhances its aggregation into globular oligomers with HTTNT cores and exposed polyQ. In a second step, a new, amyloid-like aggregate is formed with a core composed of both HTTNT and polyQ. The results indicate unprecedented complexity in how primary sequence controls aggregation within a substantially disordered peptide and have implications for the molecular mechanism of Huntington's disease.

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Figure 1: Aggregation kinetics of huntingtin exon 1 mimic peptides exploring various polyQ repeat lengths.
Figure 2: Electron micrographs of various HTTNT-related aggregates.
Figure 3: State of expansion of the HTTNT peptide in solution.
Figure 4: Concentration-dependent CD spectra of HTTNT.
Figure 5: Proton NMR analysis of HTTNT.
Figure 6: PONDR analysis of the first 600 amino acids of the human huntingtin sequence.
Figure 7: Time course of aggregation of HTTNTQ30P6 (F17W) by multiple analyses.
Figure 8: Time course of aggregation of HTTNTQ20P10 by multiple analyses.
Figure 9: Mechanism of HTTNT-mediated exon 1 aggregation.

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Acknowledgements

The authors acknowledge J. Ko and P. Patterson (California Institute of Technology) for the gift of the MW1 antibody, and T. Fullam (Allegheny College) for providing a set of aggregation kinetics data. We also acknowledge the following funding sources that contributed to the work described here: NIH R01 AG019322 (R.W.); Huntington's Disease Society of America postdoctoral fellowship (V.M.C.); NSF MCB-0444049 (T.P.C.); Petroleum Research Fund/American Chemical Society 43138-AC4 (T.P.C.); grant #4100026429 from the Commonwealth of Pennsylvania (A.M.G.).

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  1. Ashwani K Thakur and Murali Jayaraman: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Structural Biology,

    Ashwani K Thakur, Murali Jayaraman, Rakesh Mishra, Monika Thakur, In-Ja L Byeon, Dalaver H Anjum, Ravindra Kodali, James F Conway, Angela M Gronenborn & Ronald Wetzel

  2. Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh School of Medicine, Pittsburgh, 15260, Pennsylvania, USA

    Ashwani K Thakur, Murali Jayaraman, Rakesh Mishra, Monika Thakur, Ravindra Kodali & Ronald Wetzel

  3. Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, 40536, Kentucky, USA

    Veronique M Chellgren & Trevor P Creamer

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Thakur, A., Jayaraman, M., Mishra, R. et al. Polyglutamine disruption of the huntingtin exon 1 N terminus triggers a complex aggregation mechanism. Nat Struct Mol Biol 16, 380–389 (2009). https://doi.org/10.1038/nsmb.1570

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