The chloroplast 32 kDa protein is synthesized on thylakoid-bound ribosomes in Chlamydomonas reinhardtii

A cloned cpDNA fragment containing a portion of the gene for the 32–36 kDa thylakoid protein of Chlamydomonas (polypeptide D-1) was isolated. Hybridization probing of RNA from soluble and membrane fractions of Chlamydomonas showed that the mRNA for D-1 is bound to thylakoid membranes. Run-off translation of thylakoid-bound polysomes (rough thylakoids) with [³⁵S]-methionine yields polypeptide D-1 as the major product. Peptide mapping with S. aureus V-8 protease of D-1 synthesized (1) in vivo, (2) in vitro by rough thylakoids and (3) in the reticulocyte lysate directed by non-polyadenylated RNA showed that D-1 is synthesized as a precursor in the reticulocyte lysate but as the mature polypeptide by rough thylakoids.