The Hsp70-interacting protein, Hip, cooperates with the chaperone Hsp70 in protein folding and prevention of aggregation. Hsp70 interacts with non-native protein substrates in an ATP-dependent reaction cycle regulated by J-domain proteins and nucleotide exchange factors (NEFs). Hip is thought to delay substrate release by slowing ADP dissociation from Hsp70. Here we present crystal structures of the dimerization domain and the tetratricopeptide repeat (TPR) domain of rat Hip. As shown in a cocrystal structure, the TPR core of Hip interacts with the Hsp70 ATPase domain through an extensive interface, to form a bracket that locks ADP in the binding cleft. Hip and NEF binding to Hsp70 are mutually exclusive, and thus Hip attenuates active cycling of Hsp70–substrate complexes. This mechanism explains how Hip enhances aggregation prevention by Hsp70 and facilitates transfer of specific proteins to downstream chaperones or the proteasome.
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The authors thank M.B. Cox (University of Texas, El Paso, Texas, USA) for providing the S. cerevisiae DSY-1100 strain, and the MPIB core and crystallization facilities for assistance with CD spectrometry and crystallization screening. Crystal diffraction and SAXS experiments were performed at the European Synchrotron Radiation Facility (ESRF), Grenoble, France. We are grateful to D. Makino for collecting diffraction data and to C. Basquin for help with SAXS data analysis. Support by the Joint Structural Biology Group staff at ESRF Grenoble is gratefully acknowledged.
The authors declare no competing financial interests.
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Li, Z., Hartl, F. & Bracher, A. Structure and function of Hip, an attenuator of the Hsp70 chaperone cycle. Nat Struct Mol Biol 20, 929–935 (2013). https://doi.org/10.1038/nsmb.2608
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