MRNAs for two ribosomal proteins are preferentially translated in the chloroplast of Chlamydomonas reinhardtii under conditions of reduced protein synthesis

Abstract

Two mutants of the green alga Chlamydomonas reinhardtii, one deficient in the small subunit of the chloroplast ribosome and the other having chloroplast ribosomes with reduced function under certain conditions, show a characteristic syndrome of photosynthetic defects resulting from reduced chloroplast protein synthesis. These include subnormal levels of ribulose 1,5-bisphosphate carboxylase (Rubisco), reduced Hill reaction activity, diminished capacity to fix CO2, and abnormal thylakoid stacking. However, these mutants accumulate normal appearing chloroplast ribosome monomers or large subunits containing normal ribosomal protein components. In this paper, we demonstrate that pulse-labeled cells of these mutants synthesize two large subunit chloroplast ribosomal proteins at about 60% of the wild-type rate, whereas Rubisco large subunit (LSU) and the alpha subunit of CF1 are made at only 4 to 8% of the wild-type rate. No difference in the rate of turnover between ribosomal proteins and Rubisco LSU in mutant and wild-type cells was observed during a subsequent 60 min chase. Differences between the mutants and wild-type cells in the relative synthesis rates of these proteins were not reflected in the relative levels of mRNA (either hybridizable or in vitro translatable). In aggregate, these data suggest that C. reinhardtii preferentially translates chloroplast ribosomal protein mRNAs under conditions of reduced total chloroplast protein synthesis.