Dataset Information

Visualization of the Cdc48 AAA+ ATPase protein unfolding pathway: Substrate Identification

ABSTRACT: The Cdc48 AAA+ ATPase is an abundant and essential enzyme that unfolds substrates in multiple protein quality control pathways. The enzyme includes two conserved AAA+ ATPase cassettes, D1 and D2, that assemble as hexameric rings with D1 stacked above D2. Here, we report an ensemble of structures of Cdc48 affinity purified from lysate in complex with the adaptor Shp1 in the act of unfolding substrate. Our analysis reveals a continuum of structural snapshots that spans the entire translocation cycle. These data reveal new elements of Shp1-Cdc48 binding and support a “hand-over-hand” mechanism in which the sequential movement of individual subunits is closely coordinated. D1 hydrolyzes ATP and disengages from substrate prior to D2, while D2 rebinds ATP and re-engages with substrate prior to D1, thereby explaining the dominant role played by D2 in substrate translocation/unfolding.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: John Price

LAB HEAD: John C. Price

PROVIDER: PXD048280 | Pride | 2024-10-17

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	ATPhydrolysisProteinAnalysisRC3101uRC3133.csv	Csv
	PeterShen_S10_ATP_Elution_Shp1_0days_rep1O1.raw	Raw
	PeterShen_S11_ADP_Hydro_0days_rep1O1.raw	Raw
	PeterShen_S12_ADP_Hydro_2_0days_rep1O1.raw	Raw
	PeterShen_S27_4-15_Shp1_Hydro_Rep1_0days_rep1O1.raw	Raw

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Publications

Visualization of the Cdc48 AAA+ ATPase protein unfolding pathway.

Cooney Ian I Schubert Heidi L HL Cedeno Karina K Fisher Olivia N ON Carson Richard R Price John C JC Hill Christopher P CP Shen Peter S PS

Nature communications 20240829 1

The Cdc48 AAA+ ATPase is an abundant and essential enzyme that unfolds substrates in multiple protein quality control pathways. The enzyme includes two conserved AAA+ ATPase motor domains, D1 and D2, that assemble as hexameric rings with D1 stacked above D2. Here, we report an ensemble of native structures of Cdc48 affinity purified from budding yeast lysate in complex with the adaptor Shp1 in the act of unfolding substrate. Our analysis reveals a continuum of structural snapshots that spans the ...[more]

PMID: 39209885

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