RAF2 is a RuBisCO assembly factor in Arabidopsis thaliana

Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) catalyzes the reaction between gaseous carbon dioxide (CO2) and ribulose-1,5-bisphosphate. Although it is one of the most studied enzymes, the assembly mechanisms of the large hexadecameric RuBisCO is still emerging. In bacteria and in the C4 plant Zea mays, a protein with distant homology to pterin-4 alpha-carbinolamine dehydratase(PCD) has recently been shown to be involved in RuBisCO assembly. However, studies of the homologous PCD-like protein (RAF2, RuBisCO assembly factor 2) in the C3 plant Arabidopsis thaliana (A. thaliana) have so far focused on its role in hormone and stress signaling. We investigated whether A. thalianaRAF2 is also involved in RuBisCO assembly. We localized RAF2 to the soluble chloroplast stroma and demonstrated that raf2 A. thaliana mutant plants display a severe pale green phenotype with reduced levels of stromal RuBisCO. We concluded that the RAF2 protein is probably involved in RuBisCO assembly in the C3 plant A. thaliana. Significance Statement RAF2 is a factor involved in Rubisco assembly in C4 plants, but surprisingly, in the C3 plant Arabidopsis mediates Abscisic acid-dependent stress response. Here we not only determine the subcellular localization and expression pattern of RAF2, but also show by both invitro and invivo measurements that RAF2 is involved in Rubisco assembly in the C3 plant Arabidopsis. Our results shed light for further investigations on the proposed unique dual function of RAF2.