title: The chloroplast small heat-shock protein oligomer is not phosphorylated and does not dissociate during heat stress in vivo.
abstract: Plants synthesize several classes of small (15- to 30-kD monomer) heat-shock proteins (sHSPs) in response to heat stress, including a nuclear-encoded, chloroplast-localized sHSP (HSP21). Cytosolic sHSPs exist as large oligomers (approximately 200-800 kD) composed solely or primarily of sHSPs. Phosphorylation of mammalian sHSPs causes oligomer dissociation, which appears to be important for regulation of sHSP function. We examined the native structure and phosphorylation of chloroplast HSP21 to understand this protein's basic properties and to compare it with cytosolic sHSPs. The apparent size of native HSP21 complexes was > 200 kD and they did not dissociate during heat stress. We found no evidence that HSP21 or the plant cytosolic sHSPs are phosphorylated in vivo. A partial HSP21 complex purified from heat-stressed pea (Pisum sativum L.) leaves contained no proteins other than HSP21. Mature recombinant pea and Arabidopsis thaliana HSP21 were expressed in Escherichia coli, and purified recombinant Arabidopsis HSP21 assembled into homo-oligomeric complexes with the same apparent molecular mass as HSP21 complexes observed in heat-stressed leaf tissue. We propose that the native, functional form of chloroplast HSP21 is a large, oligomeric complex containing nine or more HSP21 subunits, and that plant sHSPs are not regulated by phosphorylation-induced dissociation.
authors.affiliation: Department of Biochemistry, University of Arizona, Tucson, Arizona 85721-0106, USA.
authors.forename: T C
authors.initials: TC
authors.lastname: Suzuki
authors.forename: D C
authors.initials: DC
authors.lastname: Krawitz
authors.forename: E
authors.initials: E
authors.lastname: Vierling
mesh_terms: D017360:Arabidopsis; D002736:Chloroplasts / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D018869:Heat-Shock Response / Q000502:physiology*; D006358:Hot Temperature; D046911:Macromolecular Substances; D018532:Pisum sativum; D010766:Phosphorylation; D018515:Plant Leaves / Q000378:metabolism; D010940:Plant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D011108:Polymers
chemical_list: D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D010940:Plant Proteins; D011108:Polymers