title: Rubisco, rubisco activase and ribulose-5-phosphate kinase gene expression and polypeptide accumulation in a tobacco mutant defective in chloroplast protein synthesis.
abstract: Expression of the genes for ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; rbcS and rbcL), Rubisco activase (rca) and ribulose-5-phosphate (Ru5-P) kinase (prk) and accumulation of the polypeptides was examined in chlorophyllous and chlorotic sectors of the DP1 mutant of Nicotiana tabacum. Plastids from chlorotic sectors of this variegated plastome mutant contained 30S and 50S ribosomal subunits, but had abnormally low levels of plastid polysomes. Consequently, mutant plastids were translationally repressed, unable to synthesize plastid-encoded polypeptides including the large subunit of Rubisco despite the presence of the corresponding mRNAs. Transcripts of rbcS accumulated to near wild type levels in chlorotic sectors, but there was little accumulation of the Rubisco small subunit (SS) polypeptide or holoenzyme. Messenger-RNA isolated from chlorotic sectors effectively directed the synthesis of Rubisco SS in vitro suggesting that posttranslational factors were responsible for the decrease in Rubisco SS abundance. Transcripts of rca and prk also accumulated to near wild type levels in chlorotic sectors and a diurnal rhythm in the abundance of rca mRNA was detected in green and chlorotic sectors. Despite the low abundance of Rubisco holoenzyme in chlorotic sectors, Rubisco activase and Ru5-P kinase polypeptides accumulated to significant levels. Activities of Rubisco and Ru5-P kinase paralleled protein levels, indicating that active forms of these enzymes were present in chlorotic sectors. The data indicate that the developmental events governing the accumulation of Rubisco activase and Ru5-P kinase polypeptides and the diurnal regulation of rca expression were not dependent on the attainment of photosynthetically competent plastids or the accumulation of Rubisco.
authors.affiliation: US Department of Agriculture, Agricultural Research Service and Agronomy Department, University of Kentucky, 40546-0076, Lexington, KY, USA.
authors.forename: R R
authors.initials: RR
authors.lastname: Klein
authors.forename: M E
authors.initials: ME
authors.lastname: Salvucci