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TitleStructural basis of ClpXP recognition and unfolding of ssrA-tagged substrates.
Journal, issue, pagesElife, Vol. 9, Year 2020
Publish dateOct 22, 2020
AuthorsXue Fei / Tristan A Bell / Sarah R Barkow / Tania A Baker / Robert T Sauer /
PubMed AbstractWhen ribosomes fail to complete normal translation, all cells have mechanisms to ensure degradation of the resulting partial proteins to safeguard proteome integrity. In and other eubacteria, the ...When ribosomes fail to complete normal translation, all cells have mechanisms to ensure degradation of the resulting partial proteins to safeguard proteome integrity. In and other eubacteria, the tmRNA system rescues stalled ribosomes and adds an ssrA tag or degron to the C-terminus of the incomplete protein, which directs degradation by the AAA+ ClpXP protease. Here, we present cryo-EM structures of ClpXP bound to the ssrA degron. C-terminal residues of the ssrA degron initially bind in the top of an otherwise closed ClpX axial channel and subsequently move deeper into an open channel. For short-degron protein substrates, we show that unfolding can occur directly from the initial closed-channel complex. For longer degron substrates, our studies illuminate how ClpXP transitions from specific recognition into a nonspecific unfolding and translocation machine. Many AAA+ proteases and protein-remodeling motors are likely to employ similar multistep recognition and engagement strategies.
External linksElife / PubMed:33089779 / PubMed Central
MethodsEM (single particle)
Resolution2.88 - 3.16 Å
Structure data

21875

6wr2

EMDB-21875, PDB-6wr2:
ClpP and ClpX IGF loop in ClpX-ClpP complex bound to ssrA tagged GFP
Method: EM (single particle) / Resolution: 2.88 Å

21882

6wrf

EMDB-21882, PDB-6wrf:
ClpX-ClpP complex bound to GFP-ssrA, recognition complex
Method: EM (single particle) / Resolution: 3.14 Å

21892

6wsg

EMDB-21892, PDB-6wsg:
ClpX-ClpP complex bound to ssrA-tagged GFP, intermediate complex
Method: EM (single particle) / Resolution: 3.16 Å

Chemicals

ChemComp-HOH:
WATER / Water

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM

ChemComp-MG:
Unknown entry

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

Source
  • escherichia coli (E. coli)
  • escherichia coli (strain k12) (bacteria)
  • aequorea victoria (jellyfish)
  • synthetic construct (others)
KeywordsCHAPERONE / Protein degradation / AAA+ protease complex

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