Structural insights into cyanobacterial RuBisCO assembly coordinated by two chaperones Raf1 and RbcX

RuBisCO is the most abundant enzyme in nature, catalyzing the fixation of CO2 in photosynthesis. Its common form consists of eight RbcL and eight RbcS subunits, the assembly of which requires a series of chaperones that include RbcX and RuBisCO accumulation factor 1 (Raf1). To understand how these RuBisCO-specific chaperones function during cyanobacterial RbcL(8)RbcS(8) (L8S8) holoenzyme formation, we solved a 3.3-angstrom cryo-electron microscopy structure of a 32-subunit RbcL(8)Raf1(8)RbcX(16) (L8F8X16) assembly intermediate from Anabaena sp. PCC 7120. Comparison to the previously resolved L8F8 and L8X16 structures together with biochemical assays revealed that the L8F8X16 complex forms a rather dynamic structural intermediate, favoring RbcS displacement of Raf1 and RbcX. In vitro assays further demonstrated that both Raf1 and RbcX function to regulate RuBisCO condensate formation by restricting CcmM35 binding to the stably assembled L8S8 holoenzymes. Combined with previous findings, we propose a model on how Raf1 and RbcX work in concert to facilitate, and regulate, cyanobacterial RuBisCO assembly as well as disassembly of RuBisCO condensates.

Automated summary

This study elucidated the structure of an assembly intermediate of RuBisCO enzyme in cyanobacteria and discovered that the proteins Raf1 and RbcX facilitate and regulate RuBisCO assembly by controlling the formation of RuBisCO condensates.