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 inCell Vol. 119; no. 5; pp. 653 - 665
Main Authors Weibezahn, Jimena, Tessarz, Peter, Schlieker, Christian, Zahn, Regina, Maglica, Zeljka, Lee, Sukyeong, Zentgraf, Hanswalter, Weber-Ban, Eilika U., Dougan, David A., Tsai, Francis T.F., Mogk, Axel, Bukau, Bernd
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 24.11.2004
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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.
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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Snippet Cell survival under severe thermal stress requires the activity of the ClpB (Hsp104) AAA+ chaperone that solubilizes and reactivates aggregated proteins in...
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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
URI https://dx.doi.org/10.1016/j.cell.2004.11.027
https://www.ncbi.nlm.nih.gov/pubmed/15550247
https://search.proquest.com/docview/67096679
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