Dynamic and static components power unfolding in topologically closed rings of a AAA+ proteolytic machine

Glynn, Steven E.; Nager, Andrew R.; Baker, Tania A.; Sauer, Robert T.

doi:10.1038/nsmb.2288

Cited by 57 publications

(82 citation statements)

References 42 publications

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“…However, the defect was partially compensated by forming disulfide bond. Such compensation was also found in a similar experiment performed using ClpX, the other Clp/ Hsp100 protein (41).…”

Section: Resultssupporting

confidence: 54%

Analysis of the Cooperative ATPase Cycle of the AAA+ Chaperone ClpB from Thermus thermophilus by Using Ordered Heterohexamers with an Alternating Subunit Arrangement

Yamasaki

Oohata²,

Nakamura³

et al. 2015

Journal of Biological Chemistry

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Background: Ring-shaped ClpB hexamer hydrolyzes ATP and reactivates protein aggregates. Results: Intercalation of ATPase defective subunits into the hexamer every other subunit hampered its ATPase and disaggregation activities. Conclusion: ClpB cooperatively hydrolyzes ATP, and this cooperativity is crucial for protein disaggregation. Significance: This study presents a new method to study the cooperativity of homo-oligomeric proteins and provides insights into the common mechanism of AAAϩ ATPases.

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“…However, the defect was partially compensated by forming disulfide bond. Such compensation was also found in a similar experiment performed using ClpX, the other Clp/ Hsp100 protein (41).…”

Section: Resultssupporting

confidence: 54%

Analysis of the Cooperative ATPase Cycle of the AAA+ Chaperone ClpB from Thermus thermophilus by Using Ordered Heterohexamers with an Alternating Subunit Arrangement

Yamasaki

Oohata²,

Nakamura³

et al. 2015

Journal of Biological Chemistry

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“…Proteins were concentrated by Amicon centrifugation devices (100-kDa cutoff; Millipore) and further purified by size-exclusion chromatography on a Superdex S-200 column (GE Healthcare) in 20 mM Hepes-KOH (pH 7.5), 50 mM NaCl, 1 mM DTT, and 1 mM EDTA. SF GFP-ssrA and Kaede-ssrA were expressed and purified as described (27,28). All proteins were frozen in liquid nitrogen and stored at −80°C.…”

Section: Methodsmentioning

confidence: 99%

Architecture and assembly of the archaeal Cdc48⋅20S proteasome

Barthelme¹,

Chen²,

Grabenstatter

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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ATP-dependent proteases maintain protein quality control and regulate diverse intracellular functions. Proteasomes are primarily responsible for these tasks in the archaeal and eukaryotic domains of life. Even the simplest of these proteases function as large complexes, consisting of the 20S peptidase, a barrel-like structure composed of four heptameric rings, and one or two AAA+ (ATPase associated with a variety of cellular activities) ring hexamers, which use cycles of ATP binding and hydrolysis to unfold and translocate substrates into the 20S proteolytic chamber. Understanding how the AAA+ and 20S components of these enzymes interact and collaborate to execute protein degradation is important, but the highly dynamic nature of prokaryotic proteasomes has hampered structural characterization. Here, we use electron microscopy to determine the architecture of an archaeal Cdc48·20S proteasome, which we stabilized by site-specific cross-linking. This complex displays coaxial alignment of Cdc48 and 20S and is enzymatically active, demonstrating that AAA+ unfoldase wobbling with respect to 20S is not required for function. In the complex, the N-terminal domain of Cdc48, which regulates ATP hydrolysis and degradation, packs against the D1 ring of Cdc48 in a coplanar fashion, constraining mechanisms by which the N-terminal domain alters 20S affinity and degradation activity.AAA+ protease | dynamic wobbling model | p97/VCP P roteasomes are large macromolecular complexes that degrade misfolded or damaged proteins to maintain cellular homeostasis and quality control in all domains of life. In addition, selective proteasomal turnover of regulatory proteins is often a critical element of signaling cascades that allow cells to respond to changing conditions and environmental stress (1, 2). Proteasomes consist of the self-compartmentalized 20S peptidase and one or two AAA+ (ATPase associated with a variety of cellular activities) family ring hexamers. The α 7 β 7 β 7 α 7 ring topology of the 20S enzyme ensures that the proteolytic active sites, which reside in the β-subunits, are sequestered and can only cleave substrates that enter the chamber through narrow axial pores formed by the α-subunits (3). As a consequence, ATP-dependent unfolding of natively folded protein substrates by the AAA+ ring and subsequent polypeptide translocation through a narrow axial channel and into the 20S chamber are required for degradation (4).Although the 20S peptidase is the degradation module in all proteasomes, the associated AAA+ unfolding machines differ. For example, homohexamers of Mpa/Arc in actinobacteria and PAN in archaea serve as proteasomal motors, whereas a heterohexameric Rpt 1-6 ring in the 19S particle is the motor of the eukaryotic 26S proteasome (2, 5). Characteristic of type-I AAA+ enzymes, Mpa/Arc, PAN, and Rpt 1-6 subunits contain a single AAA+ module for ATP binding and hydrolysis (6). By contrast, as found in type-II AAA+ enzymes, the homohexameric Cdc48 motor in the recently discovered archaeal Cdc48·20S proteasome cont...

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“…For some AAA proteins, the small subdomain changes its orientation relative to the large subdomain of the same subunit during the working cycle while maintaining its interaction with the large subdomain of the neighboring subunit (31,(35)(36)(37)(38). This interaction correlates with intersubunit binding strength and the ATPase activity of AAA proteins.…”

Section: Cefigl-1-aaa Is a Hexameric Protein With Strong Atpasementioning

confidence: 99%

Structural Insights into the Unusually Strong ATPase Activity of the AAA Domain of the Caenorhabditis elegans Fidgetin-like 1 (FIGL-1) Protein

Peng

Lin

Lü

et al. 2013

Journal of Biological Chemistry

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Background:Little is known about the structure and activity of the FIGL-1 protein, whose homolog fidgetin is involved in various developmental processes in mammals. Results:The Caenorhabditis elegans FIGL-1 AAA domain has strong charge interaction between subunits. Conclusion: The strong ATPase activity of the C. elegans FIGL-1 AAA domain is contributed by its strong intersubunit interaction. Significance: These unique properties suggest that the C. elegans FIGL-1 protein possesses a distinct biological function.

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Dynamic and static components power unfolding in topologically closed rings of a AAA+ proteolytic machine

Cited by 57 publications

References 42 publications

Analysis of the Cooperative ATPase Cycle of the AAA+ Chaperone ClpB from Thermus thermophilus by Using Ordered Heterohexamers with an Alternating Subunit Arrangement

Analysis of the Cooperative ATPase Cycle of the AAA+ Chaperone ClpB from Thermus thermophilus by Using Ordered Heterohexamers with an Alternating Subunit Arrangement

Architecture and assembly of the archaeal Cdc48⋅20S proteasome

Structural Insights into the Unusually Strong ATPase Activity of the AAA Domain of the Caenorhabditis elegans Fidgetin-like 1 (FIGL-1) Protein

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