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The Hsp70 Ssz1 modulates the function of the ribosome-associated J-protein Zuo1

Nature Structural & Molecular Biology volume 12, pages 497–504 (2005)Cite this article

Abstract

J-proteins are obligate partners of Hsp70s, forming a ubiquitous class of molecular chaperone machinery. The ribosome-associated Hsp70 of yeast Ssb binds nascent polypeptides as they exit the ribosome. Here we report that the ribosome-associated J-protein Zuo1 is the partner of Ssb. However, Zuo1 efficiently stimulates the ATPase activity of Ssb only when in complex with another Hsp70, Ssz1. Ssz1 binds ATP, but none of the 11 different amino acid substitutions in the ATP-binding cleft affected Ssz1 function in vivo, suggesting that neither nucleotide binding nor hydrolysis is required. We propose that Ssz1's predominant function in the cell is to facilitate Zuo1's ability to function as a J-protein partner of Ssb on the ribosome, serving as an example of an Hsp70 family member that has evolved to carry out functions distinct from that of a chaperone.

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Figure 1: RAC, but not Zuo1, catalytically stimulates Ssb's ATPase activity.
Figure 2: H128Q-RAC fails to stimulate the ATPase activity of Ssb.
Figure 3: RAC does not stimulate Ssa1; Ssb1 is not stimulated by Ydj1 and Sis1.
Figure 4: SSB1 ATPase domain mutants rescue the phenotype of zuo1-H128Q.
Figure 5: Ssz1 binds ATP, but the interaction is unstable.
Figure 6: Effect of Ssz1's nucleotide-binding state on RAC's stimulation of Ssb.
Figure 7: Alteration of ATP-binding cleft of Ssz1 has no obvious effect on growth.

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Acknowledgements

We thank C. Pfund for generating the Ssb1 ATPase domain library and helpful comments on the manuscript, H. Hundley for initial characterization of several SSZ1 mutants, N. Lopez for providing Ydj1p and Sis1p, P. D'Silva for help in AMP-PNP purification, and S. Wilbanks for input on mutagenesis of Ssz1's ATPase domain. This work was supported by US National Institutes of Health grant GM-31107 (E.A.C.).

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Authors and Affiliations

  1. Department of Biochemistry, University of Wisconsin, Madison, 53706, Wisconsin, USA

    Peggy Huang, William Walter & Elizabeth A Craig

  2. Graduate Program in Biomolecular Chemistry, University of Wisconsin, Madison, 53706, Wisconsin, USA

    Peggy Huang

  3. Institut für Biochemie, Schafmattstrasse 18, ETH-Hönggerberg, HPME 9.1, Zürich, CH-8093, Switzerland

    Matthias Gautschi

  4. Institut für Biochemie und Molekularbiologie, Universität Freiburg, Hermann-Herder-Strasse 7, Freiburg, D-79104, Germany

    Sabine Rospert

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Correspondence to Elizabeth A Craig.

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Huang, P., Gautschi, M., Walter, W. et al. The Hsp70 Ssz1 modulates the function of the ribosome-associated J-protein Zuo1. Nat Struct Mol Biol 12, 497–504 (2005). https://doi.org/10.1038/nsmb942

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