Abstract
The heat shock protein 70 (Hsp70, human HSPA1A) plays indispensable roles in cellular stress responses and protein quality control (PQC). In the framework of PQC, it cooperates with the ubiquitin-proteasome system (UPS) to clear damaged and dysfunctional proteins in the cell. Moreover, Hsp70 itself is rapidly degraded following the recovery from stress. It was demonstrated that its fast turnover is mediated via ubiquitination and subsequent degradation by the 26S proteasome. At the same time, the effect of Hsp70 on the functional state of proteasomes has been insufficiently investigated. Here, we characterized the direct effect of recombinant Hsp70 on the activity of 20S and 26S proteasomes and studied Hsp70 degradation by the 20S proteasome in vitro. We have shown that the activity of purified 20S proteasomes is decreased following incubation with recombinant human Hsp70. On the other hand, high concentrations of Hsp70 activated 26S proteasomes. Finally, we obtained evidence that in addition to previously reported ubiquitin-dependent degradation, Hsp70 could be cleaved independent of ubiquitination by the 20S proteasome. The results obtained reveal novel aspects of the interplay between Hsp70 and proteasomes.
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Acknowledgements
Authors would like to thank Dr. Vladimir Morozov for important suggestions and fruitful discussions. We are grateful to Dr. Marina Serebryakova for the assistance with MALDI MS. MALDI MS facility became available to us in the framework of the Moscow State University Development Program PNG 5.13.
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The investigation of proteasome activity in the culture cells and extracts was supported by Ministry of Education and Science of Russian Federation (agreement no. 14.Z50.31.0014) Russian Science Foundation grant (14-50-00060) and Russian President Foundation grant (МК-3613.2017.4) to A.M.
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Morozov, A.V., Astakhova, T.M., Garbuz, D.G. et al. Interplay between recombinant Hsp70 and proteasomes: proteasome activity modulation and ubiquitin-independent cleavage of Hsp70. Cell Stress and Chaperones 22, 687–697 (2017). https://doi.org/10.1007/s12192-017-0792-y
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DOI: https://doi.org/10.1007/s12192-017-0792-y