Robust Sequence Determinants of α-Synuclein Toxicity in Yeast Implicate Membrane Binding
Robert W. Newberry,
George C. Hartoularos,
Alison M. Maxwell,
Zun Zar Chi Naing,
Nishith R. Reddy,
Daniel M. C. Schwarz,
Douglas R. Wassarman,
Taia S. Wu,
Taylor B. Cavazos,
Laurel S Estes,
Elissa A. Fink,
Miriam A. Goldman,
Yessica K. Gomez,
M. Grace Gordon,
Laura M. Gunsalus,
Matthew Camp Johnson,
Matthew G Jones,
Andrew F. Kung,
Kyle E Lopez,
Elizabeth E. McCarthy,
Lakshmi E. Miller-Vedam,
Erik J. Navarro,
Christina A. Stephens,
Stephanie A. Wankowicz,
Daniel R. Wong,
Eric D. Chow,
William F. DeGrado,
Posted 03 May 2020
bioRxiv DOI: 10.1101/2020.05.01.072884 (published DOI: 10.1021/acschembio.0c00339)
Posted 03 May 2020
Protein conformations are shaped by cellular environments, but how environmental changes alter the conformational landscapes of specific proteins in vivo remains largely uncharacterized, in part due to the challenge of probing protein structures in living cells. Here, we use deep mutational scanning to investigate how a toxic conformation of α-synuclein, a dynamic protein linked to Parkinson’s disease, responds to perturbations of cellular proteostasis. In the context of a course for graduate students in the UCSF Integrative Program in Quantitative Biology, we screened a comprehensive library of α-synuclein missense mutants in yeast cells treated with a variety of small molecules that perturb cellular processes linked to α-synuclein biology and pathobiology. We found that the conformation of α-synuclein previously shown to drive yeast toxicity—an extended, membrane-bound helix—is largely unaffected by these chemical perturbations, underscoring the importance of this conformational state as a driver of cellular toxicity. On the other hand, the chemical perturbations have a significant effect on the ability of mutations to suppress α-synuclein toxicity. Moreover, we find that sequence determinants of α-synuclein toxicity are well described by a simple structural model of the membrane-bound helix. This model predicts that α-synuclein penetrates the membrane to constant depth across its length but that membrane affinity decreases toward the C terminus, which is consistent with orthogonal biophysical measurements. Finally, we discuss how parallelized chemical genetics experiments can provide a robust framework for inquiry-based graduate coursework. ### Competing Interest Statement The authors have declared no competing interest.
- Downloaded 366 times
- Download rankings, all-time:
- Site-wide: 49,887 out of 100,621
- In biochemistry: 1,545 out of 3,441
- Year to date:
- Site-wide: 11,026 out of 100,621
- Since beginning of last month:
- Site-wide: 26,421 out of 100,621
Downloads over time
Distribution of downloads per paper, site-wide
- 20 Oct 2020: Support for sorting preprints using Twitter activity has been removed, at least temporarily, until a new source of social media activity data becomes available.
- 18 Dec 2019: We're pleased to announce PanLingua, a new tool that enables you to search for machine-translated bioRxiv preprints using more than 100 different languages.
- 21 May 2019: PLOS Biology has published a community page about Rxivist.org and its design.
- 10 May 2019: The paper analyzing the Rxivist dataset has been published at eLife.
- 1 Mar 2019: We now have summary statistics about bioRxiv downloads and submissions.
- 8 Feb 2019: Data from Altmetric is now available on the Rxivist details page for every preprint. Look for the "donut" under the download metrics.
- 30 Jan 2019: preLights has featured the Rxivist preprint and written about our findings.
- 22 Jan 2019: Nature just published an article about Rxivist and our data.
- 13 Jan 2019: The Rxivist preprint is live!