The Chlamydomonas reinhardtii proteins Ccp1 and Ccp2 are required for long-term growth, but are not necessary for efficient photosynthesis, in a low-CO2 environment

Pollock, Steve V.; Prout, Davey L.; Godfrey, Ashley C.; Lemaire, Stéphane D.; Moroney, James V.

doi:10.1007/s11103-004-2650-4

Cited by 65 publications

(46 citation statements)

References 26 publications

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“…Although the products of several limiting-CO 2 -inducible genes have been identified as putative Ci transporters on the chloroplast envelope, including LCIA, LCI1, CCP1/2, and Ycf10, or on the plasma membrane, including HLA3, none of the respective gene products have been definitively determined to transport Ci species (Burow et al, 1996;Chen et al, 1997;Rolland et al, 1997;Im and Grossman, 2001;Miura et al, 2004;Pollock et al, 2004;Mariscal et al, 2006), leaving Ci transport largely a mystery.…”

Section: Discussionmentioning

confidence: 99%

Thylakoid Lumen Carbonic Anhydrase (CAH3) Mutation Suppresses Air-Dier Phenotype of LCIB Mutant in Chlamydomonas reinhardtii

2008

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An active CO 2 -concentrating mechanism is induced when Chlamydomonas reinhardtii acclimates to limiting inorganic carbon (Ci), either low-CO 2 (L-CO 2 ; air level; approximately 0.04% CO 2 ) or very low-CO 2 (VL-CO 2 ; approximately 0.01% CO 2 ) conditions. A mutant, ad1, which is defective in the limiting-CO 2 -inducible, plastid-localized LCIB, can grow in high-CO 2 or VL-CO 2 conditions but dies in L-CO 2 , indicating a deficiency in a L-CO 2 -specific Ci uptake and accumulation system. In this study, we identified two ad1 suppressors that can grow in L-CO 2 but die in VL-CO 2 . Molecular analyses revealed that both suppressors have mutations in the CAH3 gene, which encodes a thylakoid lumen localized carbonic anhydrase. Photosynthetic rates of L-CO 2 -acclimated suppressors under acclimation CO 2 concentrations were more than 2-fold higher than ad1, apparently resulting from a more than 20-fold increase in the intracellular concentration of Ci as measured by direct Ci uptake. However, photosynthetic rates of VL-CO 2 -acclimated cells under acclimation CO 2 concentrations were too low to support growth in spite of a significantly elevated intracellular Ci concentration. We conclude that LCIB functions downstream of CAH3 in the CO 2 -concentrating mechanism and probably plays a role in trapping CO 2 released by CAH3 dehydration of accumulated Ci. Apparently dehydration by the chloroplast stromal carbonic anhydrase CAH6 of the very high internal Ci caused by the defect in CAH3 provides Rubisco sufficient CO 2 to support growth in L-CO 2 -acclimated cells, but not in VL-CO 2 -acclimated cells, even in the absence of LCIB.CO 2 serves both as the substrate for photosynthesis and as an important signal to regulate plant growth and development, so variable CO 2 concentrations can impact photosynthesis, growth, and productivity of plants. Terrestrial C 4 plants have developed a CO 2 -concentrating mechanism (CCM) involving anatomical and biochemical adaptations to accumulate a higher concentration of CO 2 as substrate Rubisco and to suppress oxygenation of ribulose-1,5-bisP, a wasteful side reaction. In contrast, a different type of CCM is induced in the unicellular green microalga Chlamydomonas reinhardtii when the supply of dissolved inorganic carbon (Ci; CO 2 and HCO 3 2

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Section: Discussionmentioning

confidence: 99%

Thylakoid Lumen Carbonic Anhydrase (CAH3) Mutation Suppresses Air-Dier Phenotype of LCIB Mutant in Chlamydomonas reinhardtii

2008

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“…Before transformation, we treated cells with gametic autolysin for 15-20 min to dissociate cell clumps. Transformation was carried out by electroporation as described with modifications (Pollock et al, 2004). Cells were re-suspended in TAP liquid medium containing 60 mM sorbitol, and then transferred to a 4 mm electroporation cuvette.…”

Section: Rescue Of Ift57-1mentioning

confidence: 99%

IFT57 stabilizes the assembled intraflagellar transport complex and mediates transport of motility-related flagellar cargo

Jiang

Hernandez

et al. 2017

Journal of Cell Science

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Intraflagellar transport (IFT) is essential for the assembly and maintenance of flagella and cilia. Recent biochemical studies have shown that IFT complex B (IFT-B) is comprised of two subcomplexes, IFT-B1 and IFT-B2. The IFT-B2 subunit IFT57 lies at the interface between IFT-B1 and IFT-B2. Here, using a Chlamydomonas reinhardtii mutant for IFT57, we tested whether IFT57 is required for IFT-B complex assembly by bridging IFT-B1 and IFT-B2 together. In the ift57-1 mutant, levels of IFT57 and other IFT-B proteins were greatly reduced at the whole-cell level. However, strikingly, in the protease-free flagellar compartment, while the level of IFT57 was reduced, the levels of other IFT particle proteins were not concomitantly reduced but were present at the wild-type level. The IFT movement of the IFT57-deficient IFT particles was also unchanged. Moreover, IFT57 depletion disrupted the flagellar waveform, leading to cell swimming defects. Analysis of the mutant flagellar protein composition showed that certain axonemal proteins were altered. Taken together, these findings suggest that IFT57 does not play an essential structural role in the IFT particle complex but rather functions to prevent it from degradation. Additionally, IFT57 is involved in transporting specific motility-related proteins.

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“…To test for the ability of WT FAP46 to rescue the fap46-1 phenotype, fap46-1 (mt+) was transformed by electroporation with pFAP46. Electroporation conditions were modified from Pollock et al, (Pollock et al, 2004) as follows: Cells grown in M medium were harvested and transferred to 1/3 volume of TAP medium for 4 hr. The cells were then concentrated to 2X10 8 cells ml 21 in TAP + 40 mM sucrose; 80 ml of cells + 1 mg EcoRI-linearized pFAP46 were pulsed once in a 0.1 cm cuvette with an ECM 600 electroporator (BTX) at 200 V (2000 V cm…”

Section: Insertional Mutagenesis and Identification Of Insert Boundariesmentioning

confidence: 99%

A FAP46 mutant provides new insights into the function and assembly of the C1d complex of the ciliary central apparatus

Brown¹,

DiPetrillo²,

Smith³

et al. 2012

Journal of Cell Science

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SummaryVirtually all motile eukaryotic cilia and flagella have a '9+2' axoneme in which nine doublet microtubules surround two singlet microtubules. Associated with the central pair of microtubules are protein complexes that form at least seven biochemically and structurally distinct central pair projections. Analysis of mutants lacking specific projections has indicated that each may play a unique role in the control of flagellar motility. One of these is the C1d projection previously shown to contain the proteins FAP54, FAP46, FAP74 and FAP221/Pcdp1, which exhibits Ca 2+ -sensitive calmodulin binding. Here we report the isolation and characterization of a Chlamydomonas reinhardtii null mutant for FAP46. This mutant, fap46-1, lacks the C1d projection and has impaired motility, confirming the importance of this projection for normal flagellar movement. Those cells that are motile have severe defects in phototaxis and the photoshock response, underscoring a role for the C1d projection in Ca 2+ -mediated flagellar behavior. The data also reveal for the first time that the C1d projection is involved in the control of interdoublet sliding velocity. Our studies further identify a novel C1d subunit that we term C1d-87, give new insight into relationships between the C1d subunits, and provide evidence for multiple sites of calmodulin interaction within the C1d projection. These results represent significant advances in our understanding of an important but little studied axonemal structure.

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The Chlamydomonas reinhardtii proteins Ccp1 and Ccp2 are required for long-term growth, but are not necessary for efficient photosynthesis, in a low-CO2 environment

Cited by 65 publications

References 26 publications

Thylakoid Lumen Carbonic Anhydrase (CAH3) Mutation Suppresses Air-Dier Phenotype of LCIB Mutant in Chlamydomonas reinhardtii

Thylakoid Lumen Carbonic Anhydrase (CAH3) Mutation Suppresses Air-Dier Phenotype of LCIB Mutant in Chlamydomonas reinhardtii

IFT57 stabilizes the assembled intraflagellar transport complex and mediates transport of motility-related flagellar cargo

A FAP46 mutant provides new insights into the function and assembly of the C1d complex of the ciliary central apparatus

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