Rubisco Accumulation Factor 1 from <scp>T</scp>hermosynechococcus elongatus participates in the final stages of ribulose‐1,5‐bisphosphate carboxylase/oxygenase assembly in <scp>E</scp>scherichia coli cells and in vitro

Kolesinski, Piotr; Belusiak, Iwona; Czarnocki-Cieciura, Mariusz; Szczepaniak, Andrzej

doi:10.1111/febs.12928

Cited by 35 publications

(43 citation statements)

References 31 publications

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“…2) that gain entry into the chaperone might still misfold, condemning the mutant protein to a perpetual cycle of misfolding and unfolding within the chaperone until its degradation or aggregation in vivo (reviewed in Lin and Rye, 2006). Moreover, RbcL chimeras that are successfully folded by GroEL-GroES might still fail to assemble as complete holoenzymes in the absence of interaction with other Rubisco assemblyassisting proteins (Cloney et al, 1993;reviewed in Gutteridge and Gatenby, 1995;Saschenbrecker et al, 2007;Bracher et al, 2011;Kolesinski et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

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Engineering of chimeric eukaryotic/bacterial Rubisco large subunits in Escherichia coli

2016

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Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a rate-limiting photosynthetic enzyme that catalyzes carbon fixation in the Calvin cycle. Much interest has been devoted to engineering this ubiquitous enzyme with the goal of increasing plant growth. However, experiments that have successfully produced improved Rubisco variants, via directed evolution in Escherichia coli, are limited to bacterial Rubisco because the eukaryotic holoenzyme cannot be produced in E. coli. The present study attempts to determine the specific differences between bacterial and eukaryotic Rubisco large subunit primary structure that are responsible for preventing heterologous eukaryotic holoenzyme formation in E. coli. A series of chimeric Synechococcus Rubiscos were created in which different sections of the large subunit were swapped with those of the homologous Chlamydomonas Rubisco. Chimeric holoenzymes that can form in vivo would indicate that differences within the swapped sections do not disrupt holoenzyme formation. Large subunit residues 1-97, 198-247 and 448-472 were successfully swapped without inhibiting holoenzyme formation. In all ten chimeras, protein expression was observed for the separate subunits at a detectable level. As a first approximation, the regions that can tolerate swapping may be targets for future engineering.

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Section: Discussionmentioning

confidence: 99%

“…Holoenzyme formation itself is a series of steps involving post-translational protein processing, recognition-folding by host chaperones and assembly events, most of which involve a growing list of assisting-protein partners that is yet to be fully defined (Onizuka et al, 2004;Saschenbrecker et al, 2007;Kolesinski et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Engineering of chimeric eukaryotic/bacterial Rubisco large subunits in Escherichia coli

2016

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“…Results of chemical cross-linking experiments in maize leaves suggest all three proteins might associate with the S-subunit during Rubisco biogenesis (20). Other studies, however, suggest RAF1 interacts with post-CPN folded L-subunits to assist in L 2 then (L 2 ) 4 formation (19,22). This function mirrors that shown for RbcX, a Rubisco chaperone that acts as a "molecular staple" to assemble folded L-subunits into L 2 units for (L 2 ) 4 assembly before S-subunit binding to displace the RbcX and trigger catalytic potential (18).…”

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confidence: 99%

Improving recombinant Rubisco biogenesis, plant photosynthesis and growth by coexpressing its ancillary RAF1 chaperone

Whitney

Birch

Kelso

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

114

100

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Enabling improvements to crop yield and resource use by enhancing the catalysis of the photosynthetic CO 2 -fixing enzyme Rubisco has been a longstanding challenge. Efforts toward realization of this goal have been greatly assisted by advances in understanding the complexities of Rubisco's biogenesis in plastids and the development of tailored chloroplast transformation tools. Here we generate transplastomic tobacco genotypes expressing Arabidopsis Rubisco large subunits (AtL), both on their own (producing tob AtL plants) and with a cognate Rubisco accumulation factor 1 (AtRAF1) chaperone (producing tob plants) that has undergone parallel functional coevolution with AtL. We show AtRAF1 assembles as a dimer and is produced in tob and Arabidopsis leaves at 10-15 nmol AtRAF1 monomers per square meter. Consistent with a postchaperonin large (L)-subunit assembly role, the AtRAF1 facilitated two to threefold improvements in the amount and biogenesis rate of hybrid L 8 A S 8 t Rubisco [comprising AtL and tobacco small (S) subunits] in tob AtL-R1 leaves compared with tob AtL , despite >threefold lower steady-state Rubisco mRNA levels in tob . Accompanying twofold increases in photosynthetic CO 2 -assimilation rate and plant growth were measured for tob lines. These findings highlight the importance of ancillary protein complementarity during Rubisco biogenesis in plastids, the possible constraints this has imposed on Rubisco adaptive evolution, and the likely need for such interaction specificity to be considered when optimizing recombinant Rubisco bioengineering in plants.T he increasing global demands for food supply, bioenergy production, and CO 2 -sequestration have placed a high need on improving agriculture yields and resource use (1, 2). It is now widely recognized that yield increases are possible by enhancing the light harvesting and CO 2 -fixation processes of photosynthesis (3-5). A major target for improvement is the enzyme Rubisco [ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase] whose deficiencies in CO 2 -fixing speed and efficiency pose a key limitation to photosynthetic CO 2 capture (6, 7). In plants, the complex, multistep catalytic mechanism of Rubisco to bind its 5-carbon substrate RuBP, orient its C-2 for carboxylation, and then process the 6-carbon product into two 3-phosphoglycerate (3PGA) products, limits its throughput to one to four catalytic cycles per second (8). The mechanism also makes Rubisco prone to competitive inhibition by O 2 that produces only one 3PGA and 2-phosphoglycolate (2PG). Metabolic recycling of 2PG by photorespiration requires energy and results in most plants losing 30% of their fixed CO 2 (5). To compensate for these catalytic limitations, plants like rice and wheat invest up to 50% of the leaf protein into Rubisco, which accounts for ∼25% of their leaf nitrogen (9).Natural diversity in Rubisco catalysis demonstrates that plant Rubisco is not the pinnacle of evolution (6, 7). Better-performing versions in some red algae have the potential to raise the yield of ...

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“…Indeed, a recent screen of photosynthetic mutants in maize identified a nuclearencoded chloroplast protein, Raf1 (Rubisco accumulation factor 1), that is required for efficient Rubisco biogenesis 24 . Raf1 is conserved in all photosynthetic organisms containing RbcX and functions in Rubisco assembly in vitro and in vivo 25,26 .Here we set out to functionally and structurally characterize the plant and cyanobacterial Raf1 proteins. We solved the crystal structures of the A. thaliana Raf1 domains and analyzed the interaction of Raf1 with RbcL by multiple biochemical and biophysical approaches.…”

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confidence: 99%

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Structure and mechanism of the Rubisco-assembly chaperone Raf1

Hauser

Bhat

Miličić

et al. 2015

Nat Struct Mol Biol

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Biogenesis of the photosynthetic enzyme Rubisco, a complex of eight large (RbcL) and eight small (RbcS) subunits, requires assembly chaperones. Here we analyzed the role of Rubisco accumulation factor1 (Raf1), a dimer of ∼40-kDa subunits. We find that Raf1 from Synechococcus elongatus acts downstream of chaperonin-assisted RbcL folding by stabilizing RbcL antiparallel dimers for assembly into RbcL8 complexes with four Raf1 dimers bound. Raf1 displacement by RbcS results in holoenzyme formation. Crystal structures show that Raf1 from Arabidopsis thaliana consists of a β-sheet dimerization domain and a flexibly linked α-helical domain. Chemical cross-linking and EM reconstruction indicate that the β-domains bind along the equator of each RbcL2 unit, and the α-helical domains embrace the top and bottom edges of RbcL2. Raf1 fulfills a role similar to that of the assembly chaperone RbcX, thus suggesting that functionally redundant factors ensure efficient Rubisco biogenesis.

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Rubisco Accumulation Factor 1 from Thermosynechococcus elongatus participates in the final stages of ribulose‐1,5‐bisphosphate carboxylase/oxygenase assembly in Escherichia coli cells and in vitro

Cited by 35 publications

References 31 publications

Engineering of chimeric eukaryotic/bacterial Rubisco large subunits in <i>Escherichia coli</i>

Engineering of chimeric eukaryotic/bacterial Rubisco large subunits in <i>Escherichia coli</i>

Improving recombinant Rubisco biogenesis, plant photosynthesis and growth by coexpressing its ancillary RAF1 chaperone

Structure and mechanism of the Rubisco-assembly chaperone Raf1

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