An inorganic carbon transport system responsible for acclimation specific to air levels of CO2 in Chlamydomonas reinhardtii.
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The air-dier pmp1 and ad1 mutations lie in LciB
"Molecular analyses revealed that the Ad1/Pmp1 protein is encoded by LciB, a gene previously identified as a CO(2)-responsive gene."
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lcib mutants die at air CO2 but grow at high and very low CO2
"we identified and characterized a pmp1 allelic mutant, air dier 1 (ad1) that, like pmp1, cannot grow in low CO(2) (350 ppm) but can grow either in high CO(2) (5% CO(2)) or in very low CO(2) (<200 ppm)."
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LCIB is one of four related soluble plastid proteins
"LciB and three related genes in C. reinhardtii compose a unique gene family that encode four closely related, apparently soluble plastid proteins with no clearly identifiable conserved motifs."
Light and low-CO2-dependent LCIB-LCIC complex localization in the chloroplast supports the carbon-concentrating mechanism in Chlamydomonas reinhardtii.
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LCIB interacts with LCIC in yeast two-hybrid and in vivo
"we provide evidence to demonstrate that LCIB interacts with the LCIB homologous protein LCIC in yeast and in vivo."
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LCIB and LCIC co-localize around the pyrenoid at low CO2 in the light and form a ~350 kDa hexamer
"We also show that LCIB and LCIC are co-localized in the vicinity of the pyrenoid under LC conditions in the light, forming a hexamer complex of approximately 350 kDa, as estimated by gel filtration chromatography."
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LCIB diffuses away from the pyrenoid when the CCM is inactive
"In contrast, in the dark or under high-CO2 conditions when the CCM was inactive, LCIB immediately diffused away from the pyrenoid."
Structural insights into the LCIB protein family reveals a new group of β-carbonic anhydrases.
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Chlamydomonas LCIB and LCIC crystal structures have beta-CA-like active-site motifs
"We determined the crystal structures of LCIB and limiting CO2-inducible C protein (LCIC) from C. reinhardtii and a CA-functional homolog from Phaeodactylum tricornutum, all of which harbor motifs bearing close resemblance to the active site of canonical β-CAs."
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Three of six LCIB-family homologs tested are active CAs (per the full text these were two diatom and one bacterial homolog, not the Chlamydomonas proteins)
"Three of the six homologs are functional carbonic anhydrases (CAs)."
LCIB functions as a carbonic anhydrase: evidence from yeast and Arabidopsis carbonic anhydrase knockout mutants.
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CA activity of LCIB/LCIC had not been demonstrated before this study
"Although both LCIB and LCIC are structurally similar to βCAs, their CA activity has not been demonstrated to date."
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LCIB complements yeast NCE103 and Arabidopsis betaCA5 knockout mutants
"We provide evidence that LCIB is an active CA using a Saccharomyces cerevisiae CA knockout mutant (∆NCE103) and an Arabidopsis thaliana βCA5 knockout mutant (βca5)."
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Truncated LCIB rescues yeast; full-length LCIB rescues Arabidopsis betaca5 at low CO2
"We show that different truncated versions of the LCIB protein complement ∆NCE103, while the full length LCIB protein complements βca5 plants, so that both the yeast and plant mutants can grow in low CO2 conditions."
Functional integration of LCIB, a component of the algal carbon-concentrating mechanism, enhances carbon assimilation, nitrogen-use efficiency, and biomass in tobacco.
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LCIB expressed in tobacco chloroplast stroma raises total leaf CA activity about threefold
"LCIB was exclusively localized in the stroma and remained biologically active, leading to a ~3-fold increase in total carbonic anhydrase activity and a lower apparent CO2 compensation point (-10%)."
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Two independent lines show higher CA activity in leaf extracts
"Leaf extracts from LB-3 and LB-6 lines showed significantly higher total carbonic anhydrase activity than wild-type and vector controls."
Thylakoid lumen carbonic anhydrase (CAH3) mutation suppresses air-Dier phenotype of LCIB mutant in Chlamydomonas reinhardtii.
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cah3 mutations suppress the lcib air-dier phenotype, placing LCIB downstream of CAH3 in trapping released CO2
"We conclude that LCIB functions downstream of CAH3 in the CO2-concentrating mechanism and probably plays a role in trapping CO2 released by CAH3 dehydration of accumulated Ci."
Insertional suppressors of Chlamydomonas reinhardtii that restore growth of air-dier lcib mutants in low CO2.
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LCIB is proposed to trap CO2 released by CAH3
"is proposed to play a role in trapping CO(2) released by CAH3 (thylakoid lumen carbonic anhydrase) catalyzed dehydration of accumulated Ci, especially in low CO(2) (L-CO(2); ~0.04% CO(2)) conditions."
LCIB in the Chlamydomonas CO2-concentrating mechanism.
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Review (pre-structure) describing LCIB as a key CCM component of undefined biochemical function
"LCIB, which has been demonstrated as a key player in the eukaryotic algal CO2-concentrating mechanism (CCM), is a novel protein in Chlamydomonas lacking any recognizable domain or motif, and its exact function in the CCM has not been clearly defined."
Acclimation to very low CO2: contribution of limiting CO2 inducible proteins, LCIB and LCIA, to inorganic carbon uptake in Chlamydomonas reinhardtii.
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LCIB acts in CO2 uptake, distinct from LCIA bicarbonate transport; both needed at very low CO2
"Although both LCIA and LCIB are essential for very low CO2 acclimation, LCIB appears to function in a CO2 uptake system, whereas LCIA appears to be associated with a HCO3(-) transport system."
Knockdown of limiting-CO2-induced gene HLA3 decreases HCO3- transport and photosynthetic Ci affinity in Chlamydomonas reinhardtii.
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LCIB has a role in chloroplast Ci accumulation, partly overlapping with HLA3 and LCIA systems
"Collectively, the data presented here provide compelling evidence that HLA3 is directly or indirectly involved in HCO 3 − transport, along with additional evidence supporting a role for LCIA in chloroplast envelope HCO 3 − transport and a role for LCIB in chloroplast Ci accumulation."
CO2-dependent migration and relocation of LCIB, a pyrenoid-peripheral protein in Chlamydomonas reinhardtii.
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LCIB is in the chloroplast stroma at high and low CO2
"LC-inducible protein B (LCIB), structurally characterized as carbonic anhydrase, localizes in the chloroplast stroma under CO2-supplied and LC conditions."
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At very low CO2 LCIB aggregates around the pyrenoid
"In VLC conditions, it migrates to aggregate around the pyrenoid, where the CO2-fixing enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase is enriched."
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Relocation requires LCIC and is switched at ~7 uM CO2
"These results suggest that the localization changes of LCIB require LCIC and are controlled by CO2 concentration with ∼7 µM as the boundary."
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LCIC is lost when LCIB is disrupted
"In B3 cells, LCIC (49 kDa), the interacting protein of LCIB (48 kDa), was also hardly detected as with the previously reported LCIB RNAi strains"
Pyrenoid Starch Sheath Is Required for LCIB Localization and the CO(2)-Concentrating Mechanism in Green Algae.
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Starch sheath is required for LCIB peripheral localization
"These results suggest that the starch sheath around the pyrenoid is required for the correct localization of LCIB and for the operation of CCM."
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LCIB and LCIC are soluble stromal proteins without starch-binding domains
"Because LCIB and LCIC do not have a starch-binding domain and exist as soluble proteins in the chloroplast stroma"
Isolation and characterization of mutants defective in the localization of LCIB, an essential factor for the carbon-concentrating mechanism in Chlamydomonas reinhardtii.
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Mutants with mislocalized LCIB have reduced Ci affinity and CO2 fixation
"abl-1 and abl-3 with dispersed and speckled localization of LCIB in the chloroplast showed significant decreases in Ci affinity, Ci accumulation, and CO2 fixation."
Modelling the pyrenoid-based CO(2)-concentrating mechanism provides insights into its operating principles and a roadmap for its engineering into crops.
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Model of LCIB/LCIC as stromal CA recapturing CO2 at the pyrenoid periphery
"Consequently, LCIB relocalized near the starch sheath increases energy efficiency by recapturing CO2 molecules that diffuse out of the matrix and trapping them as HCO3− in the chloroplast"
A Spatial Interactome Reveals the Protein Organization of the Algal CO(2)-Concentrating Mechanism.
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LCIB and LCIC appear together with EPYC1 in the CCM spatial interactome
"EPYC1, LCIB and LCIC (Figure 5F)."
Expression profiling-based identification of CO2-responsive genes regulated by CCM1 controlling a carbon-concentrating mechanism in Chlamydomonas reinhardtii.