Heat shock protein 90 (HSP90) contributes to cytosolic translocation of extracellular antigen for cross-presentation by dendritic cells

In antigen (Ag) cross-presentation, dendritic cells (DCs) take up extracellular Ag and translocate them from the endosome to the cytosol for proteasomal degradation. The processed peptides can enter the conventional MHC I pathway. The molecules responsible for the translocation of Ag across the endosomal membrane into the cytosol are unknown. Here we demonstrate that heat shock protein 90 (HSP90) is critical for this step. Cross-presentation and -priming were decreased in both HSP90α-null DCs and mice. CD8α+ DC apoptosis mediated by translocation of exogenous cytochrome c to the cytosol was also eliminated in HSP90α-null mice. Ag translocation into the cytosol was diminished in HSP90α-null DCs and in DCs treated with an HSP90 inhibitor. Internalized Ag was associated with HSP90 and translocated to the cytosol, a process abrogated by the HSP90 inhibitor. Ag within purified phagosomes was released in an HSP90-dependent manner. These results demonstrate the important role of HSP90 in cross-presentation by pulling endosomal Ag out into the cytosol.

[1]  L. Huber,et al.  Biogenesis of phagolysosomes proceeds through a sequential series of interactions with the endocytic apparatus , 1994, The Journal of cell biology.

[2]  Keiji Tanaka,et al.  Two Distinct Pathways Mediated by PA28 and hsp90 in Major Histocompatibility Complex Class I Antigen Processing , 2002, The Journal of experimental medicine.

[3]  A. Goldberg,et al.  Degradation of cell proteins and the generation of MHC class I-presented peptides. , 1999, Annual review of immunology.

[4]  J. Yewdell,et al.  The exception that reinforces the rule: crosspriming by cytosolic peptides that escape degradation. , 2008, Immunity.

[5]  J. Davoust,et al.  ER–phagosome fusion defines an MHC class I cross-presentation compartment in dendritic cells , 2003, Nature.

[6]  Etienne Gagnon,et al.  Endoplasmic Reticulum-Mediated Phagocytosis Is a Mechanism of Entry into Macrophages , 2002, Cell.

[7]  P. Srivastava,et al.  Peptides chaperoned by heat-shock proteins are a necessary and sufficient source of antigen in the cross-priming of CD8+ T cells , 2005, Nature Immunology.

[8]  L. Neckers,et al.  The Heat Shock Protein 90 Antagonist Novobiocin Interacts with a Previously Unrecognized ATP-binding Domain in the Carboxyl Terminus of the Chaperone* , 2000, The Journal of Biological Chemistry.

[9]  M. McComb,et al.  The cytosolic entry of diphtheria toxin catalytic domain requires a host cell cytosolic translocation factor complex , 2003, Journal of Cell Biology.

[10]  P. Srivastava,et al.  Isolation of MHC class I-restricted tumor antigen peptide and its precursors associated with heat shock proteins hsp70, hsp90, and gp96. , 1999, Journal of immunology.

[11]  W. Heath,et al.  Cross-presentation, dendritic cells, tolerance and immunity. , 2001, Annual review of immunology.

[12]  P. Cresswell,et al.  A role for the endoplasmic reticulum protein retrotranslocation machinery during crosspresentation by dendritic cells. , 2006, Immunity.

[13]  Keiji Tanaka,et al.  Hsp90-mediated Assembly of the 26 S Proteasome Is Involved in Major Histocompatibility Complex Class I Antigen Processing* , 2008, Journal of Biological Chemistry.

[14]  J. Blum,et al.  HSP90α and HSP90β Isoforms Selectively Modulate MHC Class II Antigen Presentation in B Cells1 , 2009, The Journal of Immunology.

[15]  T. Rapoport,et al.  Polyubiquitination is required for US11-dependent movement of MHC class I heavy chain from endoplasmic reticulum into cytosol. , 2001, Molecular biology of the cell.

[16]  Etienne Gagnon,et al.  Phagosomes are competent organelles for antigen cross-presentation , 2003, Nature.

[17]  A. Iwasaki,et al.  Autophagy-Dependent Viral Recognition by Plasmacytoid Dendritic Cells , 2007, Science.

[18]  D. Mok,et al.  Modulation of Chaperone Function and Cochaperone Interaction by Novobiocin in the C-terminal Domain of Hsp90 , 2006, Journal of Biological Chemistry.

[19]  J. Miller,et al.  Cross-presentation: a general mechanism for CTL immunity and tolerance. , 1998, Immunology today.

[20]  K. Teter,et al.  Hsp90 Is Required for Transfer of the Cholera Toxin A1 Subunit from the Endoplasmic Reticulum to the Cytosol* , 2010, The Journal of Biological Chemistry.

[21]  Zihai Li,et al.  Cutting Edge: Cross-Presentation of Cell-Associated Antigens to MHC Class I Molecule Is Regulated by a Major Transcription Factor for Heat Shock Proteins1 , 2004, The Journal of Immunology.

[22]  P. Workman.  Altered states: selectively drugging the Hsp90 cancer chaperone. , 2004, Trends in molecular medicine.

[23]  Nicholas J. Hoogenraad,et al.  Molecular Chaperones Hsp90 and Hsp70 Deliver Preproteins to the Mitochondrial Import Receptor Tom70 , 2003, Cell.

[24]  T. Nakayama,et al.  Essential Role of Endogenous Heat Shock Protein 90 of Dendritic Cells in Antigen Cross-Presentation , 2010, The Journal of Immunology.

[25]  P. Srivastava,et al.  Heat-shock protein 90 associates with N-terminal extended peptides and is required for direct and indirect antigen presentation , 2008, Proceedings of the National Academy of Sciences.

[26]  Dirk Tiemann,et al.  The Host Cell Chaperone Hsp90 Is Essential for Translocation of the Binary Clostridium botulinum C2 Toxin into the Cytosol* , 2003, Journal of Biological Chemistry.

[27]  C. Taxis,et al.  Protein dislocation from the ER requires polyubiquitination and the AAA-ATPase Cdc48 , 2002, Nature Cell Biology.

[28]  J. Miyazaki,et al.  A transgenic mouse line that retains Cre recombinase activity in mature oocytes irrespective of the cre transgene transmission. , 1997, Biochemical and biophysical research communications.

[29]  N. Shastri,et al.  Hsp90alpha chaperones large C-terminally extended proteolytic intermediates in the MHC class I antigen processing pathway. , 2006, Immunity.

[30]  Li Wu,et al.  Selective suicide of cross-presenting CD8+ dendritic cells by cytochrome c injection shows functional heterogeneity within this subset , 2008, Proceedings of the National Academy of Sciences.

[31]  E. Unanue,et al.  Batf3 Deficiency Reveals a Critical Role for CD8α+ Dendritic Cells in Cytotoxic T Cell Immunity , 2008, Science.

[32]  T. Rapoport,et al.  Retro-translocation of proteins from the endoplasmic reticulum into the cytosol , 2002, Nature Reviews Molecular Cell Biology.

[33]  Peter Claus,et al.  FGF-1 and FGF-2 Require the Cytosolic Chaperone Hsp90 for Translocation into the Cytosol and the Cell Nucleus* , 2006, Journal of Biological Chemistry.

[34]  L. Neckers,et al.  Extracellular heat shock protein 90: A role for a molecular chaperone in cell motility and cancer metastasis , 2007, Cancer science.

[35]  L. Neckers,et al.  A small molecule cell-impermeant Hsp90 antagonist inhibits tumor cell motility and invasion , 2008, Oncogene.

[36]  Kristin A. Hogquist,et al.  T cell receptor antagonist peptides induce positive selection , 1994, Cell.

[37]  Anna M. Keller,et al.  Identification of a dendritic cell receptor that couples sensing of necrosis to immunity , 2009, Nature.

[38]  J. Neefjes,et al.  Export of antigenic peptides from the endoplasmic reticulum intersects with retrograde protein translocation through the Sec61p channel. , 2000, Immunity.

[39]  S. Tonegawa,et al.  TAP1 mutant mice are deficient in antigen presentation, surface class I molecules, and CD4−8+ T cells , 1992, Cell.

[40]  M. de Virgilio,et al.  Ubiquitination Is Required for the Retro-translocation of a Short-lived Luminal Endoplasmic Reticulum Glycoprotein to the Cytosol for Degradation by the Proteasome* , 1998, The Journal of Biological Chemistry.