DnaK Functions as a Central Hub in the E. coli Chaperone Network

SummaryCellular chaperone networks prevent potentially toxic protein aggregation and ensure proteome integrity. Here, we used Escherichia coli as a model to understand the organization of these networks, focusing on the cooperation of the DnaK system with the upstream chaperone Trigger factor (TF) and the downstream GroEL. Quantitative proteomics revealed that DnaK interacts with at least ∼700 mostly cytosolic proteins, including ∼180 relatively aggregation-prone proteins that utilize DnaK extensively during and after initial folding. Upon deletion of TF, DnaK interacts increasingly with ribosomal and other small, basic proteins, while its association with large multidomain proteins is reduced. DnaK also functions prominently in stabilizing proteins for subsequent folding by GroEL. These proteins accumulate on DnaK upon GroEL depletion and are then degraded, thus defining DnaK as a central organizer of the chaperone network. Combined loss of DnaK and TF causes proteostasis collapse with disruption of GroEL function, defective ribosomal biogenesis, and extensive aggregation of large proteins.

Graphical AbstractFigure optionsDownload as PowerPoint slideHighlights
► The DnaK (Hsp70) chaperone interactome comprises ∼700–1000 different proteins
► About 180 proteins are highly enriched on DnaK and are relatively aggregation-prone
► Loss of Trigger factor or GroEL markedly alters the DnaK interactome
► Loss of DnaK and Trigger factor disrupts proteostasis and protein flux to GroEL