Physiological and Molecular Biological Characterization of Intracellular Carbonic Anhydrase from the Marine Diatom<i>Phaeodactylum tricornutum</i>

Satoh, Dan; Hiraoka, Yasutaka; Colman, Brian; Matsuda, Yusuke

doi:10.1104/pp.126.4.1459

Cited by 85 publications

(98 citation statements)

References 62 publications

(33 reference statements)

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“…1B, subsection a) or the coding region of mptca1 (accession no. AF414191; Satoh et al, 2001) with a deleted termination codon inserted between pre138 and egfp (Fig. 1B, b).…”

Section: Vector Constructsmentioning

confidence: 99%

“…The gene for the presequence of PtCA1 precursor, pre138 ( Fig. 1A; Satoh et al, 2001; accession no. AF414191), was ligated directly with egfp (Fig.…”

Section: Vector Constructsmentioning

confidence: 99%

“…Most of the diatoms studied so far exhibit an efficient direct uptake of HCO 3 2 from the bulk medium even in the presence of the weakly permeable CA inhibitor, acetazolamide (Badger et al, 1998). In P. tricornutum UTEX642, at least one major soluble CA (PtCA1) activity was detected and 2 mM acetazolamide were shown to be ineffective in inhibiting direct HCO 3 2 uptake, whereas 0.5 mM ethoxyzolamide, a highly permeable CA inhibitor, dramatically suppressed high-affinity photosynthesis (Matsuda et al, 2001a;Satoh et al, 2001). Considering the fact that internal forms of CAs seem to be more crucial for cell growth under CO 2 limitation than external forms in the green alga C. reinhardtii, the importance of internal CA for the primary production under CO 2 limitation would also be of significant interest in marine diatoms.…”

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confidence: 99%

“…Both cDNA (ptca1) and PtCA1 protein have been isolated and PtCA1 was shown to be a b-type CA with a well-conserved zinc coordination domain ). The ptca1 cDNA was also shown to possess a presequence of 138 bp (pre138), which encodes 46 N-terminal amino acids (Pre46AA) that do not occur in the mature PtCA1 (corresponding gene sequence is termed hereafter mptca1; Satoh et al, 2001). Pre46AA is relatively rich in hydrophobic amino acids and it has been proposed that it might function as a sorting signal for PtCA1 .…”

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confidence: 99%

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Localization of Soluble β-Carbonic Anhydrase in the Marine Diatom Phaeodactylum tricornutum. Sorting to the Chloroplast and Cluster Formation on the Girdle Lamellae

et al. 2005

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A b-carbonic anhydrase (CA) in the marine diatom Phaeodactylum tricornutum (PtCA1) is encoded by the nuclear genome. This enzyme was previously found to be important for the operation of photosynthesis with a high affinity for dissolved inorganic carbon. A cDNA sequence that encodes PtCA1 (ptca1) was shown to possess a presequence of 138 bp (pre138), which encodes an N-terminal sequence of 46 amino acids (Pre46AA) that does not exist in the mature PtCA1. In this study, pre138 was ligated with the enhanced green fluorescent protein (GFP) gene (egfp), and introduced into P. tricornutum by microprojectile bombardment. Subsequently, the expressed Pre46AA-GFP fusion was shown to be localized in the chloroplast stroma, whereas the expressed GFP without Pre46AA was localized in the cytoplasm. Insertion of the DNA sequence, encoding a mature region of ptca1 (mptca1) between pre138 and egfp, resulted in the formation of particles with concentrated GFP fluorescence in the stroma of P. tricornutum. These particles, 0.3 to 3.0 mm in size, were shown to be distinct from the mitochondria and localized on the surface of the putative girdle lamella. The attachment of the initial one-half of the pre138 to the mptca1-egfp fusion caused the expressed GFP fusion to accumulate in areas surrounding the chloroplast, presumably due to the presence of the endoplasmic reticulum signal encoded by the initial half-sequence and to the absence of the chloroplast transit sequence. These results indicate that PtCA1 is targeted to the stroma by the bipartite sequences of Pre46AA and that the observed GFP particles are formed specifically in the stroma due to the function of the mptca1.

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“…1B, subsection a) or the coding region of mptca1 (accession no. AF414191; Satoh et al, 2001) with a deleted termination codon inserted between pre138 and egfp (Fig. 1B, b).…”

Section: Vector Constructsmentioning

confidence: 99%

“…The gene for the presequence of PtCA1 precursor, pre138 ( Fig. 1A; Satoh et al, 2001; accession no. AF414191), was ligated directly with egfp (Fig.…”

Section: Vector Constructsmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Localization of Soluble β-Carbonic Anhydrase in the Marine Diatom Phaeodactylum tricornutum. Sorting to the Chloroplast and Cluster Formation on the Girdle Lamellae

et al. 2005

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“…A b-carbonic anhydrase in the marine diatom P. tricornutum (PtCA1), thought to be one of the crucial chloroplastic components of the CCM (Satoh et al, 2001;Tanaka et al, 2005), is known to be regulated strictly by the ambient [CO 2 ] and light . A promoter region of the PtCA1 gene (ptca1) was isolated previously, and it was demonstrated that the critical CO 2 -response sequence was located downstream 270 bp relative to the transcription start site .…”

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confidence: 99%

CO2 Sensing at Ocean Surface Mediated by cAMP in a Marine Diatom

et al. 2006

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Marine diatoms are known to be responsible for about a quarter of global primary production and their photosynthesis is sustained by inorganic carbon-concentrating mechanisms and/or C 4 metabolism. Activities of the inorganic carbon-concentrating mechanism are attenuated under enriched [CO 2 ]; however, impacts of this factor on primary productivity and the molecular mechanisms of CO 2 responses in marine diatoms are unknown. In this study, transgenic cells were generated of the marine diatom Phaeodactylum tricornutum by the introduction of a b-glucuronidase reporter gene under the control of an intrinsic CO 2 -responsive promoter, which is the sequence between 280 to 161 relative to the transcription start site of a chloroplasticcarbonic anhydrase gene, ptca1, obtained from P. tricornutum. The activity of the ptca1 promoter was effectively repressed in air-level CO 2 by treating cells with a 1.0 mM cAMP analog, dibutyryl cAMP, or a cAMP phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine. Deletion of the intrinsic cAMP-response element from the ptca1 promoter caused a lack of repression of the reporter gene uidA, even under elevated [CO 2 ] and a null phenotype to the strong repressive effects of dibutyryl cAMP and 3-isobutyl-1-methylxanthine on the ptca1 promoter. Deletion of the cAMP-response element was also shown to cause derepression of the uidA reporter gene in the dark. These results indicate that the cytosolic cAMP level increases under elevated [CO 2 ] and represses the ptca1 promoter. This strongly suggests the participation of cAMP metabolism, presumably at the cytosolic level, in controlling CO 2 -acquisition systems under elevated [CO 2 ] at the ocean surface in a marine diatom.Marine diatoms are responsible for one-half of primary productivity in the ocean and hence play a key role in global cycles of carbon and other inorganic nutrients (Tréguer et al., 1995;Falkowski et al., 2000).[CO 2 ] dissolved in seawater is limited under the present atmospheric pCO 2 (below 15 mM at 20°C) that is much lower than the K m [CO 2 ] of Rubisco in diatom species (Badger et al., 1998). This implies that marine diatoms need active uptake and accumulation systems for dissolved inorganic carbon (DIC) to support their photosynthesis. There is a substantial body of evidence that the operation of the inorganic carbon-concentrating mechanism (CCM) confers on marine diatom cells highaffinity photosynthesis for DIC (Colman and Rotatore, 1995;Johnston and Raven, 1996;Matsuda et al., 2001), which is due to the operation of active uptake of both CO 2 and HCO 2 3 (Colman and Rotatore, 1995;Johnston and Raven, 1996;Matsuda et al., 2001). The activity of the CCM is suppressed under CO 2 -enriched conditions, whereas it is induced in CO 2 -limiting conditions; this regulation is due to CO 2 sensing by algal cells and induction of the CCM facilitates an ample supply of CO 2 to Rubisco even under extreme [CO 2 ] limitation (Badger et al., , 1998Kaplan et al., 1980;Miller et al., 1990;Colman and Rotatore, 1995;Johnston and R...

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