Regulation of genes encoding the large subunit of ribulose-1,5-bisphosphate carboxylase and the photosystem II polypeptides D-1 and D-2 during the cell cycle of Chlamydomonas reinhardtii.

Abstract

Synthesis of the major chloroplast proteins is temporally regulated in light-dark-synchronized Chlamydomonas cells. We have used cloned chloroplast DNA probes, and in vitro and in vivo protein synthesis to examine the cell cycle regulation of photosystem II polypeptides D-1 and D-2, and the large subunit of ribulose-1,5-bisphosphate carboxylase (RuBPCase LS). Synthesis and accumulation of D-1 and D-2 mRNAs occurs during the first half of the light period (G1), correlating with increasing synthesis of the polypeptides. Rifampicin, added immediately before the light period, inhibited the normal increase in D-1, D-2 polypeptide synthesis. During the dark period D-1, D-2 mRNAs persist at high levels despite reduced rates of mRNA synthesis and translation during this period. Cell-free translation analyses indicate that the D-1 mRNA present during the dark period is efficient at directing synthesis of the D-1 precursor in vitro. We conclude that expression of the psbA (D-1) and psbD (D-2) genes are regulated primarily at the transcriptional level during the light-induction period but at the translational level for the remainder of the cell cycle. Transcripts of the RuBPCase LS gene (rbcL) are also found at high levels during the light and dark periods but, unlike D-1 and D-2, LS mRNA levels do not increase until the last half of the light period and measurable synthesis and accumulation of this mRNA occurs during the dark. Furthermore, induction of LS polypeptide synthesis during the light period is insensitive to rifampicin. We conclude that LS production is regulated primarily at the translational level during the cell cycle.