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Plant Physiology Preview Published on April 22, 2005; 10.1104/pp.104.058982
Received January 4, 2005
Localization of Soluble
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  M. Lapointe, T. D.B. MacKenzie, and D. Morse An External {delta}-Carbonic Anhydrase in a Free-Living Marine Dinoflagellate May Circumvent Diffusion-Limited Carbon Acquisition Plant Physiology, July 1, 2008; 147(3): 1427 - 1436. [Abstract] [Full Text] [PDF]
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H. Harada, K. Nakajima, K. Sakaue, and Y. Matsuda CO2 Sensing at Ocean Surface Mediated by cAMP in a Marine Diatom Plant Physiology, November 1, 2006; 142(3): 1318 - 1328. [Abstract] [Full Text] [PDF]
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 A. R. Soto, H. Zheng, D. Shoemaker, J. Rodriguez, B. A. Read, and T. M. Wahlund Identification and Preliminary Characterization of Two cDNAs Encoding Unique Carbonic Anhydrases from the Marine Alga Emiliania huxleyi Appl. Envir. Microbiol., August 1, 2006; 72(8): 5500 - 5511. [Abstract] [Full Text] [PDF]
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H. Harada, D. Nakatsuma, M. Ishida, and Y. Matsuda Regulation of the Expression of Intracellular {beta}-Carbonic Anhydrase in Response to CO2 and Light in the Marine Diatom Phaeodactylum tricornutum Plant Physiology, October 1, 2005; 139(2): 1041 - 1050. [Abstract] [Full Text] [PDF]
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-Carbonic Anhydrase in the Marine Diatom Phaeodactylum tricornutum. Sorting to the Chloroplast and Cluster Formation on the Girdle Lamellae
-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 µm 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.