Structural analysis of BAG1 cochaperone and its interactions with Hsc70 heat shock protein

Klára Briknarová; Shinichi Takayama; Lars Brive; Marnie L. Havert; Deborah A. Knee; Jesus Velasco; Sachiko Homma; Edelmira Cabezas; Joan Stuart; David W. Hoyt; Arnold C. Satterthwait; Miguel Llinás; John C. Reed; Kathryn R. Ely

doi:10.1038/86236

Structural analysis of BAG1 cochaperone and its interactions with Hsc70 heat shock protein

Klára Briknarová
, Shinichi Takayama
, Lars Brive
, Marnie L. Havert
, Deborah A. Knee
, Jesus Velasco
, Sachiko Homma
, Edelmira Cabezas
, Joan Stuart
, David W. Hoyt
, Arnold C. Satterthwait
, Miguel Llinás
, John C. Reed
, Kathryn R. Ely

Chemistry and Biochemistry

Research output: Contribution to journal › Article › peer-review

147 Scopus citations

Abstract

BAG-family proteins share a conserved protein interaction region, called the 'BAG domain' which binds and regulates Hsp70/Hsc70 molecular chaperones. This family of cochaperones functionally regulates signal transducing proteins and transcription factors important for cell stress responses, apoptosis, proliferation, cell migration and hormone action. Aberrant overexpression of the founding member of this family, BAG1, occurs in human cancers. In this study, a structure-based approach was used to identify interacting residues in a BAG1-Hsc70 complex. An Hsc70-binding fragment of BAG1 was shown by multidimensional NMR methods to consist of an antiparallel three-helix bundle. NMR chemical shift experiments marked surface residues on the second (α2) and third (α3) helices in the BAG domain that are involved in chaperone binding. Structural predictions were confirmed by site-directed mutagenesis of these residues, resulting in loss of binding of BAG1 to Hsc70 in vitro and in cells. Molecular docking of BAG1 to Hsc70 and mutagenesis of Hsc70 marked the molecular surface of the ATPase domain necessary for interaction with BAG1. The results provide a structural basis for understanding the mechanism by which BAG proteins link molecular chaperones and cell signaling pathways.

Original language	English
Pages (from-to)	349-352
Number of pages	4
Journal	Nature Structural Biology
Volume	8
Issue number	4
DOIs	https://doi.org/10.1038/86236
State	Published - 2001

Funding

This research was performed in part at the Environmental Molecular Sciences Laboratory (a national scientific user facility sponsored by the U.S. DOE Office of Biological and Environmental Research) located at Pacific Northwest National Laboratory, operated by Battelle for the DOE. The authors are grateful to the staff at the HFMR facility at EMSL for useful discussions. We also thank K. Baker for protein purification and characterization, X. Jia for assistance with the Varian 500 MHz spectrometer and P. Crescenti for manuscript preparation. This work was funded by the National Institutes of Health NCI, NIHLB, USAMRDC Breast Cancer Program, the University of California Breast Cancer Research Program, the State of California Cancer Research Program, the Susan G. Komen Breast Cancer Foundation, the Human Frontier Science Program and CaPCURE.

Funders
Pacific Northwest National Laboratory

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

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10.1038/86236

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@article{d80f3793d6614443929850eec9f546ad,

title = "Structural analysis of BAG1 cochaperone and its interactions with Hsc70 heat shock protein",

abstract = "BAG-family proteins share a conserved protein interaction region, called the 'BAG domain' which binds and regulates Hsp70/Hsc70 molecular chaperones. This family of cochaperones functionally regulates signal transducing proteins and transcription factors important for cell stress responses, apoptosis, proliferation, cell migration and hormone action. Aberrant overexpression of the founding member of this family, BAG1, occurs in human cancers. In this study, a structure-based approach was used to identify interacting residues in a BAG1-Hsc70 complex. An Hsc70-binding fragment of BAG1 was shown by multidimensional NMR methods to consist of an antiparallel three-helix bundle. NMR chemical shift experiments marked surface residues on the second (α2) and third (α3) helices in the BAG domain that are involved in chaperone binding. Structural predictions were confirmed by site-directed mutagenesis of these residues, resulting in loss of binding of BAG1 to Hsc70 in vitro and in cells. Molecular docking of BAG1 to Hsc70 and mutagenesis of Hsc70 marked the molecular surface of the ATPase domain necessary for interaction with BAG1. The results provide a structural basis for understanding the mechanism by which BAG proteins link molecular chaperones and cell signaling pathways.",

author = "Kl{\'a}ra Briknarov{\'a} and Shinichi Takayama and Lars Brive and Havert, \{Marnie L.\} and Knee, \{Deborah A.\} and Jesus Velasco and Sachiko Homma and Edelmira Cabezas and Joan Stuart and Hoyt, \{David W.\} and Satterthwait, \{Arnold C.\} and Miguel Llin{\'a}s and Reed, \{John C.\} and Ely, \{Kathryn R.\}",

note = "Funding Information: This research was performed in part at the Environmental Molecular Sciences Laboratory (a national scientific user facility sponsored by the U.S. DOE Office of Biological and Environmental Research) located at Pacific Northwest National Laboratory, operated by Battelle for the DOE. The authors are grateful to the staff at the HFMR facility at EMSL for useful discussions. We also thank K. Baker for protein purification and characterization, X. Jia for assistance with the Varian 500 MHz spectrometer and P. Crescenti for manuscript preparation. This work was funded by the National Institutes of Health NCI, NIHLB, USAMRDC Breast Cancer Program, the University of California Breast Cancer Research Program, the State of California Cancer Research Program, the Susan G. Komen Breast Cancer Foundation, the Human Frontier Science Program and CaPCURE.",

year = "2001",

doi = "10.1038/86236",

language = "English",

volume = "8",

pages = "349--352",

journal = "Nature Structural Biology",

issn = "1072-8368",

publisher = "Nature Publishing Group",

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T1 - Structural analysis of BAG1 cochaperone and its interactions with Hsc70 heat shock protein

AU - Briknarová, Klára

AU - Takayama, Shinichi

AU - Brive, Lars

AU - Havert, Marnie L.

AU - Knee, Deborah A.

AU - Velasco, Jesus

AU - Homma, Sachiko

AU - Cabezas, Edelmira

AU - Stuart, Joan

AU - Hoyt, David W.

AU - Satterthwait, Arnold C.

AU - Llinás, Miguel

AU - Reed, John C.

AU - Ely, Kathryn R.

N1 - Funding Information: This research was performed in part at the Environmental Molecular Sciences Laboratory (a national scientific user facility sponsored by the U.S. DOE Office of Biological and Environmental Research) located at Pacific Northwest National Laboratory, operated by Battelle for the DOE. The authors are grateful to the staff at the HFMR facility at EMSL for useful discussions. We also thank K. Baker for protein purification and characterization, X. Jia for assistance with the Varian 500 MHz spectrometer and P. Crescenti for manuscript preparation. This work was funded by the National Institutes of Health NCI, NIHLB, USAMRDC Breast Cancer Program, the University of California Breast Cancer Research Program, the State of California Cancer Research Program, the Susan G. Komen Breast Cancer Foundation, the Human Frontier Science Program and CaPCURE.

PY - 2001

Y1 - 2001

N2 - BAG-family proteins share a conserved protein interaction region, called the 'BAG domain' which binds and regulates Hsp70/Hsc70 molecular chaperones. This family of cochaperones functionally regulates signal transducing proteins and transcription factors important for cell stress responses, apoptosis, proliferation, cell migration and hormone action. Aberrant overexpression of the founding member of this family, BAG1, occurs in human cancers. In this study, a structure-based approach was used to identify interacting residues in a BAG1-Hsc70 complex. An Hsc70-binding fragment of BAG1 was shown by multidimensional NMR methods to consist of an antiparallel three-helix bundle. NMR chemical shift experiments marked surface residues on the second (α2) and third (α3) helices in the BAG domain that are involved in chaperone binding. Structural predictions were confirmed by site-directed mutagenesis of these residues, resulting in loss of binding of BAG1 to Hsc70 in vitro and in cells. Molecular docking of BAG1 to Hsc70 and mutagenesis of Hsc70 marked the molecular surface of the ATPase domain necessary for interaction with BAG1. The results provide a structural basis for understanding the mechanism by which BAG proteins link molecular chaperones and cell signaling pathways.

AB - BAG-family proteins share a conserved protein interaction region, called the 'BAG domain' which binds and regulates Hsp70/Hsc70 molecular chaperones. This family of cochaperones functionally regulates signal transducing proteins and transcription factors important for cell stress responses, apoptosis, proliferation, cell migration and hormone action. Aberrant overexpression of the founding member of this family, BAG1, occurs in human cancers. In this study, a structure-based approach was used to identify interacting residues in a BAG1-Hsc70 complex. An Hsc70-binding fragment of BAG1 was shown by multidimensional NMR methods to consist of an antiparallel three-helix bundle. NMR chemical shift experiments marked surface residues on the second (α2) and third (α3) helices in the BAG domain that are involved in chaperone binding. Structural predictions were confirmed by site-directed mutagenesis of these residues, resulting in loss of binding of BAG1 to Hsc70 in vitro and in cells. Molecular docking of BAG1 to Hsc70 and mutagenesis of Hsc70 marked the molecular surface of the ATPase domain necessary for interaction with BAG1. The results provide a structural basis for understanding the mechanism by which BAG proteins link molecular chaperones and cell signaling pathways.

UR - https://www.scopus.com/pages/publications/0035069393

U2 - 10.1038/86236

DO - 10.1038/86236

M3 - Article

C2 - 11276257

AN - SCOPUS:0035069393

SN - 1072-8368

VL - 8

SP - 349

EP - 352

JO - Nature Structural Biology

JF - Nature Structural Biology

IS - 4

ER -

Structural analysis of BAG1 cochaperone and its interactions with Hsc70 heat shock protein

Abstract

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UN SDGs

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