Thermotolerance Requires Refolding of Aggregated Proteins by Substrate Translocation through the Central Pore of ClpB
Cell survival under severe thermal stress requires the activity of the ClpB (Hsp104) AAA+ chaperone that solubilizes and reactivates aggregated proteins in concert with the DnaK (Hsp70) chaperone system. How protein disaggregation is achieved and whether survival is solely dependent on ClpB-mediated...
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Published in | Cell Vol. 119; no. 5; pp. 653 - 665 |
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Main Authors | , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Elsevier Inc
24.11.2004
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Subjects | |
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Abstract | Cell survival under severe thermal stress requires the activity of the ClpB (Hsp104) AAA+ chaperone that solubilizes and reactivates aggregated proteins in concert with the DnaK (Hsp70) chaperone system. How protein disaggregation is achieved and whether survival is solely dependent on ClpB-mediated elimination of aggregates or also on reactivation of aggregated proteins has been unclear. We engineered a ClpB variant, BAP, which associates with the ClpP peptidase and thereby is converted into a degrading disaggregase. BAP translocates substrates through its central pore directly into ClpP for degradation. ClpB-dependent translocation is demonstrated to be an integral part of the disaggregation mechanism. Protein disaggregation by the BAP/ClpP complex remains dependent on DnaK, defining a role for DnaK at early stages of the disaggregation reaction. The activity switch of BAP to a degrading disaggregase does not support thermotolerance development, demonstrating that cell survival during severe thermal stress requires reactivation of aggregated proteins. |
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AbstractList | Cell survival under severe thermal stress requires the activity of the ClpB (Hsp104) AAA+ chaperone that solubilizes and reactivates aggregated proteins in concert with the DnaK (Hsp70) chaperone system. How protein disaggregation is achieved and whether survival is solely dependent on ClpB-mediated elimination of aggregates or also on reactivation of aggregated proteins has been unclear. We engineered a ClpB variant, BAP, which associates with the ClpP peptidase and thereby is converted into a degrading disaggregase. BAP translocates substrates through its central pore directly into ClpP for degradation. ClpB-dependent translocation is demonstrated to be an integral part of the disaggregation mechanism. Protein disaggregation by the BAP/ClpP complex remains dependent on DnaK, defining a role for DnaK at early stages of the disaggregation reaction. The activity switch of BAP to a degrading disaggregase does not support thermotolerance development, demonstrating that cell survival during severe thermal stress requires reactivation of aggregated proteins. |
Author | Schlieker, Christian Weibezahn, Jimena Dougan, David A. Tessarz, Peter Maglica, Zeljka Zentgraf, Hanswalter Weber-Ban, Eilika U. Mogk, Axel Zahn, Regina Tsai, Francis T.F. Lee, Sukyeong Bukau, Bernd |
Author_xml | – sequence: 1 givenname: Jimena surname: Weibezahn fullname: Weibezahn, Jimena organization: Zentrum für Molekulare Biologie der Universität Heidelberg, Universität Heidelberg, Im Neuenheimer Feld 282, Heidelberg D-69120, Germany – sequence: 2 givenname: Peter surname: Tessarz fullname: Tessarz, Peter organization: Zentrum für Molekulare Biologie der Universität Heidelberg, Universität Heidelberg, Im Neuenheimer Feld 282, Heidelberg D-69120, Germany – sequence: 3 givenname: Christian surname: Schlieker fullname: Schlieker, Christian organization: Zentrum für Molekulare Biologie der Universität Heidelberg, Universität Heidelberg, Im Neuenheimer Feld 282, Heidelberg D-69120, Germany – sequence: 4 givenname: Regina surname: Zahn fullname: Zahn, Regina organization: Zentrum für Molekulare Biologie der Universität Heidelberg, Universität Heidelberg, Im Neuenheimer Feld 282, Heidelberg D-69120, Germany – sequence: 5 givenname: Zeljka surname: Maglica fullname: Maglica, Zeljka organization: Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule, Zürich, CH-8093, Switzerland – sequence: 6 givenname: Sukyeong surname: Lee fullname: Lee, Sukyeong organization: Department of Biochemistry and Molecular Biology, Baylor College of Medicine, 1 Baylor Plaza, Houston, TX 77030 USA – sequence: 7 givenname: Hanswalter surname: Zentgraf fullname: Zentgraf, Hanswalter organization: Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, Heidelberg D-69120, Germany – sequence: 8 givenname: Eilika U. surname: Weber-Ban fullname: Weber-Ban, Eilika U. organization: Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule, Zürich, CH-8093, Switzerland – sequence: 9 givenname: David A. surname: Dougan fullname: Dougan, David A. organization: Zentrum für Molekulare Biologie der Universität Heidelberg, Universität Heidelberg, Im Neuenheimer Feld 282, Heidelberg D-69120, Germany – sequence: 10 givenname: Francis T.F. surname: Tsai fullname: Tsai, Francis T.F. organization: Department of Biochemistry and Molecular Biology, Baylor College of Medicine, 1 Baylor Plaza, Houston, TX 77030 USA – sequence: 11 givenname: Axel surname: Mogk fullname: Mogk, Axel email: a.mogk@zmbh.uni-heidelberg.de organization: Zentrum für Molekulare Biologie der Universität Heidelberg, Universität Heidelberg, Im Neuenheimer Feld 282, Heidelberg D-69120, Germany – sequence: 12 givenname: Bernd surname: Bukau fullname: Bukau, Bernd email: bukau@zmbh.uni-heidelberg.de organization: Zentrum für Molekulare Biologie der Universität Heidelberg, Universität Heidelberg, Im Neuenheimer Feld 282, Heidelberg D-69120, Germany |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/15550247$$D View this record in MEDLINE/PubMed |
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SubjectTerms | Cell Survival - genetics Endopeptidase Clp Escherichia coli - genetics Escherichia coli - metabolism Escherichia coli Proteins - genetics Escherichia coli Proteins - isolation & purification Escherichia coli Proteins - metabolism Heat-Shock Proteins - genetics Heat-Shock Proteins - isolation & purification Heat-Shock Proteins - metabolism Heat-Shock Response - genetics HSP70 Heat-Shock Proteins - genetics HSP70 Heat-Shock Proteins - metabolism Molecular Chaperones - genetics Molecular Chaperones - metabolism Molecular Motor Proteins Molecular Sequence Data Multiprotein Complexes - genetics Multiprotein Complexes - metabolism Protein Engineering Protein Folding Protein Transport - physiology Sequence Homology, Amino Acid Sequence Homology, Nucleic Acid |
Title | Thermotolerance Requires Refolding of Aggregated Proteins by Substrate Translocation through the Central Pore of ClpB |
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