title: High-molecular-mass complexes formed in vivo contain smHSPs and HSP70 and display chaperone-like activity.
abstract: Stress can have profound effects on the cell. The elicitation of the stress response in the cell is often accompanied by the synthesis of high-molecular-mass complexes, sometimes termed heat shock granules (HSGs). The presence of the complexes has been shown to be important for the survival of cells subjected to stress. We purified these complexes from heat-stressed BY-2 tobacco cells. HSG complexes formed in vivo contain predominantly smHSPs, HSP40 and HSP70 and display chaperone-like activity. Tubulins as well as other proteins may be part of the complex or its substrate. The proteins, except smHSPs and to some extent HSP70, were hypersensitive to proteolysis, suggesting that they were partially denatured and not an integral part of the HSG complexes. When citrate synthase was used as the substrate, in vivo generated HSG complexes exhibited strong nucleotide-dependent in vitro chaperone activity. Measurable ATP-mediated hydrolytic activity was detected. Isolated HSG complexes are stable until ATP is added, which leads to rapid dissociation of the complex into subunits. It is proposed that smHSPs form the core of the complex in association with ATP-dependent HSP70 and HSP40 cochaperones. Implications of these findings are discussed.
authors.affiliation: Department of Plant Physiology, Charles University, Prague, Czech Republic.
authors.forename: P
authors.initials: P
authors.lastname: Smýkal
authors.forename: I
authors.initials: I
authors.lastname: Hrdý
authors.forename: P M
authors.initials: PM
authors.lastname: Pechan
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D002950:Citrate (si)-Synthase / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K / Q000378:metabolism; D004789:Enzyme Activation; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis*; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000737:chemistry; D015151:Immunoblotting; D007700:Kinetics; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D018832:Molecular Chaperones / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*; D010947:Plants, Toxic; D012542:Scattering, Radiation; D013696:Temperature; D014026:Nicotiana
chemical_list: D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D018832:Molecular Chaperones; D010940:Plant Proteins; D000255:Adenosine Triphosphate; D002950:Citrate (si)-Synthase; D019286:Endopeptidase K