The central body of the cyanelles of <i>Cyanophora paradoxa</i>: a eukaryotic carboxysome?

Abstract

The cyanelles of the glaucocystophyte Cyanophora paradoxa combine two prokaryotic features not found in other phototrophic eukaryotes: a peptidoglycan wall and a putative carboxysome. Both of them would be indispensable when a inorganic carbon concentrating mechanism involving high accumulation of bicarbonate in the cyanelle stroma is assumed. Two approaches were used. (i) An expressed sequence tag library was generated allowing access to interesting genes and microarray technology. Hybridization of the microarrays to RNA from cells grown at high and low CO 2 yielded 97 genes that were upregulated under CO 2 stress whereas 87 genes were found to be downregulated. (ii) Cyanelle central bodies were isolated and protein components other than Rubisco were investigated by mass spectrometry. So far, mass spectrometric analysis of putative carboxysomal proteins yielded only sequences with no match in the databases. Rubisco activase could be shown via in vitro import and Western blotting to be copackaged with Rubisco in isolated purified central bodies. While our data support the presence of an inorganic carbon concentrating mechanism in cyanelles, they do not allow us to distinguish the microcompartment as a carboxysome or pyrenoid.Key words: Cyanophora paradoxa, cyanelles, carboxysome, Rubisco activase, carbon-concentrating mechanism, microarrays.