Fusion protein analysis reveals the precise regulation between Hsp70 and Hsp100 during protein disaggregation

ClpB, a bacterial Hsp100, is a ring-shaped AAA+ chaperone that can reactivate aggregated proteins in cooperation with DnaK, a bacterial Hsp70, and its co-factors. ClpB subunits comprise two AAA+ modules with an interstitial rod-shaped M-domain. The M-domain regulates ClpB ATPase activity and interac...

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Bibliographic Details
Published in:Scientific reports 2017-08, Vol.7 (1), p.8648-14, Article 8648
Main Authors: Hayashi, Sayaka, Nakazaki, Yosuke, Kagii, Kei, Imamura, Hiromi, Watanabe, Yo-hei
Format: Article
Language:English
Subjects:
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Adenosine triphosphatase
ClpB protein
Cooperativity
Disaggregation
DnaK protein
Fusion protein
Hsp70 protein
Humanities and Social Sciences
multidisciplinary
Protein structure
Proteins
Science
Science (multidisciplinary)
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Summary:ClpB, a bacterial Hsp100, is a ring-shaped AAA+ chaperone that can reactivate aggregated proteins in cooperation with DnaK, a bacterial Hsp70, and its co-factors. ClpB subunits comprise two AAA+ modules with an interstitial rod-shaped M-domain. The M-domain regulates ClpB ATPase activity and interacts directly with the DnaK nucleotide-binding domain (NBD). Here, to clarify how these functions contribute to the disaggregation process, we constructed ClpB, DnaK, and aggregated YFP fusion proteins in various combinations. Notably, i) DnaK activates ClpB only when the DnaK substrate-binding domain (SBD) is in the closed conformation, affording high DnaK-peptide affinity; ii) although NBD alone can activate ClpB, SBD is required for disaggregation; and iii) tethering aggregated proteins to the activated ClpB obviates SBD requirements. These results indicate that DnaK activates ClpB only when the SBD tightly holds aggregated proteins adjacent to ClpB for effective disaggregation.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-017-08917-8