title: Transketolase from Cyanophora paradoxa: in vitro import into cyanelles and pea chloroplasts and a complex history of a gene often, but not always, transferred in the context of secondary endosymbiosis.
abstract: The glaucocystophyte Cyanophora paradoxa is an obligatorily photoautotrophic biflagellated protist containing cyanelles, peculiar plastids surrounded by a peptidoglycan layer between their inner and outer envelope membranes. Although the 136-kb cyanelle genome surpasses higher plant chloroplast genomes in coding capacity by about 50 protein genes, these primitive plastids still have to import >2,000 polypeptides across their unique organelle wall. One such protein is transketolase, an essential enzyme of the Calvin cycle. We report the sequence of the pre-transketolase cDNA from C. paradoxa and in vitro import experiments of precursor polypeptides into cyanelles and into pea chloroplasts. The transit sequence clearly indicates the localization of the gene product to cyanelles and is more similar to the transit sequences of the plant homologues than to transit sequences of other cyanelle precursor polypeptides with the exception of a cyanelle consensus sequence at the N-terminus. The mature sequence reveals conservation of the thiamine pyrophosphate binding site. A neighbor-net planar graph suggests that Cyanophora, higher plants, and the photosynthetic protist Euglena gracilis acquired their nuclear-encoded transketolase genes via endosymbiotic gene transfer from the cyanobacterial ancestor of plastids; in the case of Euglena probably entailing two transfers, once from the plastid in the green algal lineage and once again in the secondary endosymbiosis underlying the origin of Euglena's plastids. By contrast, transketolase genes in some eukaryotes with secondary plastids of red algal origin, such as Thalassiosira pseudonana, have retained the pre-existing transketolase gene germane to their secondary host.
authors.affiliation: Max F. Perutz Laboratories, Department of Biochemistry and Cell Biology, University of Vienna, Dr. Bohrgasse 9, 1030 Vienna, Austria.
authors.forename: Yan
authors.initials: Y
authors.lastname: Ma
authors.forename: Johannes
authors.initials: J
authors.lastname: Jakowitsch
authors.forename: Oliver
authors.initials: O
authors.lastname: Deusch
authors.forename: Katrin
authors.initials: K
authors.lastname: Henze
authors.forename: William
authors.initials: W
authors.lastname: Martin
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Löffelhardt
mesh_terms: D000595:Amino Acid Sequence; D002736:Chloroplasts / Q000201:enzymology*; D044424:Cyanophora / Q000201:enzymology* / Q000235:genetics; D048028:DNA, Algal / Q000032:analysis / Q000235:genetics; D005056:Euglena gracilis / Q000201:enzymology / Q000235:genetics; D022761:Gene Transfer, Horizontal; D005796:Genes; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000378:metabolism / Q000382:microbiology*; D021381:Protein Transport; D017422:Sequence Analysis, DNA; D013559:Symbiosis; D014174:Transketolase / Q000378:metabolism*
chemical_list: D048028:DNA, Algal; D014174:Transketolase