title: Intracellular localization of a group II chaperonin indicates a membrane-related function.
abstract: Chaperonins are protein complexes that are believed to function as part of a protein folding system in the cytoplasm of the cell. We observed, however, that the group II chaperonins known as rosettasomes in the hyperthermophilic archaeon Sulfolobus shibatae, are not cytoplasmic but membrane associated. This association was observed in cultures grown at 60 degrees C and 76 degrees C or heat-shocked at 85 degrees C by using immunofluorescence microscopy and in thick sections of rapidly frozen cells grown at 76 degrees C by using immunogold electron microscopy. We observed that increased abundance of rosettasomes after heat shock correlated with decreased membrane permeability at lethal temperature (92 degrees C). This change in permeability was not seen in cells heat-shocked in the presence of the amino acid analogue azetidine 2-carboxylic acid, indicating functional protein synthesis influences permeability. Azetidine experiments also indicated that observed heat-induced changes in lipid composition in S. shibatae could not account for changes in membrane permeability. Rosettasomes purified from cultures grown at 60 degrees C and 76 degrees C or heat-shocked at 85 degrees C bind to liposomes made from either the bipolar tetraether lipids of Sulfolobus or a variety of artificial lipid mixtures. The presence of rosettasomes did not significantly change the transition temperature of liposomes, as indicated by differential scanning calorimetry, or the proton permeability of liposomes, as indicated by pyranine fluorescence. We propose that these group II chaperonins function as a structural element in the natural membrane based on their intracellular location, the correlation between their functional abundance and membrane permeability, and their potential distribution on the membrane surface.
authors.affiliation: National Aeronautics and Space Administration Ames Research Center, Moffett Field, CA 94035, USA. jonathan.d.trent@nasa.gov
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Trent
authors.forename: Hiromi K
authors.initials: HK
authors.lastname: Kagawa
authors.forename: Chad D
authors.initials: CD
authors.lastname: Paavola
authors.forename: R Andrew
authors.initials: RA
authors.lastname: McMillan
authors.forename: Jeanie
authors.initials: J
authors.lastname: Howard
authors.forename: Linda
authors.initials: L
authors.lastname: Jahnke
authors.forename: Colleen
authors.initials: C
authors.lastname: Lavin
authors.forename: Tsegereda
authors.initials: T
authors.lastname: Embaye
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Henze
mesh_terms: D019843:Archaeal Proteins / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology* / Q000648:ultrastructure; D018834:Chaperonin 60 / Q000032:analysis; D018833:Chaperonins / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D005612:Freeze Drying; D056385:Group II Chaperonins; D006358:Hot Temperature; D008081:Liposomes / Q000032:analysis; D008563:Membrane Lipids / Q000032:analysis; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D017510:Protein Folding; D021381:Protein Transport; D016944:Sulfolobus / Q000502:physiology* / Q000648:ultrastructure
chemical_list: D019843:Archaeal Proteins; D018834:Chaperonin 60; D008081:Liposomes; D008563:Membrane Lipids; D018833:Chaperonins; D056385:Group II Chaperonins