Ccm1, a regulatory gene controlling the induction of a carbon-concentrating mechanism in Chlamydomonas reinhardtii by sensing CO2 availability.
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CCM1 is a nuclear-encoded 699-aa protein with an N-terminal putative zinc-finger motif and a glutamine repeat typical of transcriptional activators
"we isolated a nuclear regulatory gene, Ccm1, encoding a 699-aa hydrophilic protein with a putative zinc-finger motif in its N-terminal region and a Gln repeat characteristic of transcriptional activators"
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Ccm1 complements the C16 mutant, restoring carbon transport, pyrenoid development and induction of CCM genes
"Introduction of Ccm1 into this mutant restored an active carbon transport through the CCM, development of a pyrenoid structure in the chloroplast, and induction of a set of CCM-related genes."
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Ccm1 is essential for CO2-responsive induction of the CCM
"These data indicate that Ccm1 is essential to control the induction of CCM by sensing CO2 availability in Chlamydomonas cells."
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The cia5 mutation (His54) lies in the zinc-finger motif
"revealed that His-54 within the putative zinc-finger motif of the CCM1 is crucial to its regulatory function"
The Cia5 gene controls formation of the carbon concentrating mechanism in Chlamydomonas reinhardtii.
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The cia5 mutant fails to induce CO2-responsive transcripts and the CCM; the Cia5 gene restores the wild-type phenotype
"The cia5 mutant is defective in all of these phenotypes. A newly isolated gene, designated Cia5, restores transformed cia5 cells to the phenotype of wild-type cells."
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CIA5 mRNA and protein are constitutive in high and low CO2
"It encodes a protein that has features of a putative transcription factor and that, likewise, is present constitutively in low and high CO2 conditions."
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C-terminally truncated CIA5 causes constitutive expression of CO2-responsive genes
"cia5 mutants complemented with the truncated gene exhibit constitutive synthesis of mRNAs from CO2-responsive genes in light under both high and low CO2 conditions"
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CIA5 directly or indirectly controls transcription of CO2-responsive genes
"These discoveries suggest that posttranslational changes to the C-terminal domain control the ability of CIA5 to act as an inducer and directly or indirectly control transcription of CO2-responsive genes."
Isolation and Characterization of a Mutant of Chlamydomonas reinhardtii Deficient in the CO(2) Concentrating Mechanism.
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The original cia5 (CIA-5) mutant cannot induce the CCM or adapt to low CO2
"It is concluded that this mutant fails to induce the CO(2) concentrating system and is incapable of adapting to low CO(2) conditions."
Significance of zinc in a regulatory protein, CCM1, which regulates the carbon-concentrating mechanism in Chlamydomonas reinhardtii.
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Each of the two N-terminal regions of CCM1 binds one zinc ion, measured by atomic absorption spectrometry of GST fusions
"It was found that 1 mol of zinc is bound to 1 mol of amino acid regions 1-71 and 72-101 of CCM1, respectively."
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Zinc-coordinating Cys/His residues are required for CCM induction in vivo
"amino acid residues such as C36, C41, H54, C77, C80, H90 and C93 were indispensable for induction of the CCM in response to Ci-limiting stress conditions"
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CCM1 is part of a large complex regardless of Ci availability
"CCM1 is present as a protein complex of approximately 290-580 kDa independent of Ci availability"
Expression profiling-based identification of CO2-responsive genes regulated by CCM1 controlling a carbon-concentrating mechanism in Chlamydomonas reinhardtii.
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Induction of almost all low-CO2-inducible genes requires CCM1
"The fact that the induction of almost all low-CO2 inducible genes was impaired in the ccm1 mutant suggests that CCM1 is a master regulator of CCM through putative low-CO2 signal transduction pathways."
Transcriptome-wide changes in Chlamydomonas reinhardtii gene expression regulated by carbon dioxide and the CO2-concentrating mechanism regulator CIA5/CCM1.
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CIA5 and CO2 affect expression of almost 25% of all genes
"We observed a massive impact of CIA5 and CO 2 on the transcriptome, affecting almost 25% of all Chlamydomonas genes"
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Expression of most Ci transporters and inducible CAs requires CIA5, although CIA5 expression is CO2-independent
"Expression of most putative C i transporters and induced CAs requires CIA5, even though the expression of CIA5 itself does not depend on the CO 2 level, so posttranslational activation of CIA5 in low CO 2 apparently is required for CIA5 to regulate these genes"
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At the time, the DNA sequences recognized by CIA5 and its direct targets were unknown
"CIA5 has been proposed to be a transcription regulator ( Fukuzawa et al., 2001 ; Xiang et al., 2001 ), but we know very little about sequences recognized by its putative DNA binding domain or the genes it directly regulates downstream."
A nuclear CobW/WW-domain factor represses the CO(2)-concentrating mechanism in the green alga Chlamydomonas reinhardtii.
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CCM1-mGold expressed in the ccm1-1 mutant localizes to the nucleus under both high and very low CO2
"Confocal microscopy revealed strong nuclear signals in both transformants under HC and VLC conditions (Fig. 3A)."
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Nuclear localization of CCM1 does not change with CO2
"Notably, the nuclear localization pattern remained unchanged between HC and VLC conditions for both proteins, showing punctate nuclear signals characteristic of transcription factor localization (Fig. 3A)."
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Direct DNA binding by CCM1 had not been demonstrated
"Although CCM1 contains two Zn2+-binding sites (13, 14), direct DNA binding has not been demonstrated, suggesting that CCM1 operates in conjunction with other nuclear factors."
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CBP1 binds CCM1 constitutively in the nucleus and represses CCM1-dependent genes under high CO2
"CBP1 colocalizes and interacts with CCM1 in the nucleus regardless of CO2 conditions."
CIA5 and its interacting metal-binding GTPase ZNG3 are degraded by the proteasome in Zn deficiency.
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CIA5 protein is degraded by the proteasome under Zn deficiency, preventing CCM induction
"We demonstrate that CIA5 is constitutively expressed regardless of Zn supply, but its encoded protein undergoes proteasomal degradation under Zn deficiency, providing a checkpoint that prevents CCM induction when Zn cofactors are unavailable."
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ZNG3 is a constitutive CIA5-interacting protein
"We identified a COBW domain-containing GTPase, ZNG3, as a constitutive interacting partner of CIA5."
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CIA5 has a C2H2 zinc finger and a second zinc site in its first 110 residues
"One such example is CIA5, which contains two Zn‐binding sites within the first 110 amino acids, one in a C2H2 zinc finger between amino acids 35 and 70, and the second between amino acids 70 and 100"
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CIA5 is nuclear and required for transcriptional induction of CCM genes
"CIA5 localizes to the nucleus, and cia5 mutants fail to transcriptionally induce Ci transporter genes, CAHs, and many genes encoding structural components of the CCM"
Regulatory logic of the Chlamydomonas CO(2)-concentrating mechanism: coupling carbon flux, energy supply, and pyrenoid architecture.
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Recent in vitro work (Chen and Spalding 2026, Plant Mol Biol Rep, doi:10.1007/s11105-025-01670-7; not in PubMed) reports specific binding of purified full-length CIA5 to a GC-rich promoter motif
"Earlier studies did not detect direct DNA binding by CCM1/CIA5, but recent in vitro work using purified full-length CIA5 demonstrated specific binding to a GC-rich motif found in promoters of CIA5-dependent genes (Chen and Spalding, 2026)."
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CCM1/CIA5 appears lineage-restricted
"The Chlamydomonas transcriptional regulator CCM1/CIA5 appears to be a lineage-restricted component rather than a broadly conserved green-plant regulator."
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The upstream CO2 sensor for CIA5 is unknown
"The primary CO2/HCO3− sensor upstream of CCM1/CIA5 remains unknown."
Light-independent regulation of algal photoprotection by CO(2) availability.
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CIA5 controls LHCSR3 and PSBS transcript accumulation
"Finally, we show that the CCM regulator CIA5 also regulates photoprotection, controlling LHCSR3 and PSBS transcript accumulation while inhibiting LHCSR1 protein accumulation."
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The cia5 LHCSR3 defect is rescued by complementation
"This phenotype was fully reversed by ectopic expression of the WT CIA5 gene (Fig. 3a)."
Photosynthetic characteristics of a multicellular green alga Volvox carteri in response to external CO2 levels possibly regulated by CCM1/CIA5 ortholog.
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A Volvox ortholog conserves the zinc-binding N-terminus and putative nuclear localization/export signals
"Comparison of Volvox and Chlamydomonas CCM1 revealed a highly conserved N-terminal region containing zinc-binding amino acid residues, putative nuclear localization and export signals, and a C-terminal region containing a putative LXXLL protein-protein interaction motif."
The CO2 concentrating mechanism and photosynthetic carbon assimilation in limiting CO2 : how Chlamydomonas works against the gradient.