Robust Sequence Determinants of α-Synuclein Toxicity in Yeast Implicate Membrane Binding.

Autor:	Newberry RW; Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143, United States., Arhar T; Chemistry and Chemical Biology Graduate Program, University of California, San Francisco, California 94143, United States., Costello J; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Hartoularos GC; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Maxwell AM; Chemistry and Chemical Biology Graduate Program, University of California, San Francisco, California 94143, United States., Naing ZZC; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Pittman M; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Reddy NR; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Schwarz DMC; Chemistry and Chemical Biology Graduate Program, University of California, San Francisco, California 94143, United States., Wassarman DR; Chemistry and Chemical Biology Graduate Program, University of California, San Francisco, California 94143, United States., Wu TS; Chemistry and Chemical Biology Graduate Program, University of California, San Francisco, California 94143, United States., Barrero D; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Caggiano C; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Catching A; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Cavazos TB; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Estes LS; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Faust B; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Fink EA; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Goldman MA; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Gomez YK; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Gordon MG; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Gunsalus LM; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Hoppe N; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Jaime-Garza M; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Johnson MC; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Jones MG; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Kung AF; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Lopez KE; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Lumpe J; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Martyn C; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., McCarthy EE; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Miller-Vedam LE; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Navarro EJ; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Palar A; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Pellegrino J; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Saylor W; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Stephens CA; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Strickland J; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Torosyan H; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Wankowicz SA; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Wong DR; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Wong G; Integrative Program in Quantitative Biology, University of California, San Francisco, California 94143, United States., Redding S; Department of Biochemistry and Biophysics, University of California, San Francisco, California 94143, United States., Chow ED; Department of Biochemistry and Biophysics, University of California, San Francisco, California 94143, United States., DeGrado WF; Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143, United States., Kampmann M; Department of Biochemistry and Biophysics, University of California, San Francisco, California 94143, United States.; Institute for Neurodegenerative Disease, University of California, San Francisco, California 94143, United States.; Chan Zuckerberg Biohub, San Francisco, California 94158, United States.
Jazyk:	angličtina
Zdroj:	ACS chemical biology [ACS Chem Biol] 2020 Aug 21; Vol. 15 (8), pp. 2137-2153. Date of Electronic Publication: 2020 Aug 12.
DOI:	10.1021/acschembio.0c00339
Abstrakt:	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.
Databáze:	MEDLINE
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