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

ABSTRACT: 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 RbcL₈RbcS₈ (L₈S₈) holoenzyme formation, we solved a 3.3-Å cryo-electron microscopy structure of a 32-subunit RbcL₈Raf1₈RbcX₁₆ (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.

SUBMITTER: Li Q

PROVIDER: S-EPMC9485235 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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

Li Qiong Q Jiang Yong-Liang YL Xia Ling-Yun LY Chen Yuxing Y Zhou Cong-Zhao CZ

Cell discovery 20220920 1

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 RbcL8RbcS8 (L8S8) holoenzyme formation, we solved a 3.3-Å cryo-electron microscopy stru ...[more]

PMID: 36123352

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Project description:Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the rate-limiting step in the Calvin-Benson cycle, which transforms atmospheric carbon into a biologically useful carbon source. The slow catalytic rate of Rubisco and low substrate specificity necessitate the production of high levels of this enzyme. In order to engineer a more efficient plant Rubisco, we need to better understand its folding and assembly process. Form I Rubisco, found in green algae and vascular plants, is a hexadecamer composed of 8 large subunits (RbcL), encoded by the chloroplast genome and 8 small, nuclear-encoded subunits (RbcS). Unlike its cyanobacterial homolog, which can be reconstituted in vitro or in E. coli, assisted by bacterial chaperonins (GroEL-GroES) and the RbcX chaperone, biogenesis of functional chloroplast Rubisco requires Cpn60-Cpn20, the chloroplast homologs of GroEL-GroES, and additional auxiliary factors, including Rubisco accumulation factor 1 (Raf1), Rubisco accumulation factor 2 (Raf2) and Bundle sheath defective 2 (Bsd2). The discovery and characterization of these factors paved the way for Arabidopsis Rubisco assembly in E. coli. In the present review, we discuss the uniqueness of hetero-oligomeric chaperonin complex for RbcL folding, as well as the sequential or concurrent actions of the post-chaperonin chaperones in holoenzyme assembly. The exact stages at which each assembly factor functions are yet to be determined. Expression of Arabidopsis Rubisco in E. coli provided some insight regarding the potential roles for Raf1 and RbcX in facilitating RbcL oligomerization, for Bsd2 in stabilizing the oligomeric core prior to holoenzyme assembly, and for Raf2 in interacting with both RbcL and RbcS. In the long term, functional characterization of each known factor along with the potential discovery and characterization of additional factors will set the stage for designing more efficient plants, with a greater biomass, for use in biofuels and sustenance.

Dataset Information

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

Publications

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

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