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. 2013 Jan;30(1):70-8.
doi: 10.1093/molbev/mss142. Epub 2012 May 23.

Evolution of saxitoxin synthesis in cyanobacteria and dinoflagellates

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Evolution of saxitoxin synthesis in cyanobacteria and dinoflagellates

Jeremiah D Hackett et al. Mol Biol Evol. 2013 Jan.

Abstract

Dinoflagellates produce a variety of toxic secondary metabolites that have a significant impact on marine ecosystems and fisheries. Saxitoxin (STX), the cause of paralytic shellfish poisoning, is produced by three marine dinoflagellate genera and is also made by some freshwater cyanobacteria. Genes involved in STX synthesis have been identified in cyanobacteria but are yet to be reported in the massive genomes of dinoflagellates. We have assembled comprehensive transcriptome data sets for several STX-producing dinoflagellates and a related non-toxic species and have identified 265 putative homologs of 13 cyanobacterial STX synthesis genes, including all of the genes directly involved in toxin synthesis. Putative homologs of four proteins group closely in phylogenies with cyanobacteria and are likely the functional homologs of sxtA, sxtG, and sxtB in dinoflagellates. However, the phylogenies do not support the transfer of these genes directly between toxic cyanobacteria and dinoflagellates. SxtA is split into two proteins in the dinoflagellates corresponding to the N-terminal portion containing the methyltransferase and acyl carrier protein domains and a C-terminal portion with the aminotransferase domain. Homologs of sxtB and N-terminal sxtA are present in non-toxic strains, suggesting their functions may not be limited to saxitoxin production. Only homologs of the C-terminus of sxtA and sxtG were found exclusively in toxic strains. A more thorough survey of STX+ dinoflagellates will be needed to determine if these two genes may be specific to SXT production in dinoflagellates. The A. tamarense transcriptome does not contain homologs for the remaining STX genes. Nevertheless, we identified candidate genes with similar predicted biochemical activities that account for the missing functions. These results suggest that the STX synthesis pathway was likely assembled independently in the distantly related cyanobacteria and dinoflagellates, although using some evolutionarily related proteins. The biological role of STX is not well understood in either cyanobacteria or dinoflagellates. However, STX production in these two ecologically distinct groups of organisms suggests that this toxin confers a benefit to producers that we do not yet fully understand.

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Figures

F<sc>ig</sc>. 1.
Fig. 1.
Graph depicting the relationship between the amount of Illumina input data and the cumulative assembly length for the de novo dinoflagellate transcriptome assemblies.
F<sc>ig</sc>. 2.
Fig. 2.
(A) Partial maximum likelihood (ML) phylogeny of the N-terminal region of sxtA and candidate proteins containing the amidinotransferase domain. The full tree is in supplementary fig. S1, Supplementary Material online. For all phylogenies, numbers at the branches show the results of 100 bootstrap replicates (<50). The names at the tips indicate the taxonomy, species name, and GI or contig number of the sequences. STX-producing dinoflagellates are indicated in bold type. The (+) indicates a STX-producing organism. (B) ML phylogeny of the C-terminal region of sxtA and candidate proteins. (C) Domain structure of sxtA-related proteins in A. tamarense Group 4 compared to sxtA from Cylindrospermopsis raciborskii T3 (GI: 114462352). Protein domains were predicted using InterPro Scan. GNAT, GCN5-related N-acetyltransferase (IPR000182); PP-binding, Phosphopantetheine-binding (IPR006163); Aminotransferase 1,2, Aminotransferase class I/II (IPR004839); Nat, Acyl-CoA N-acyltransferases (IPR016181). Signal peptides were predicted by SignalP in InterproScan.
F<sc>ig</sc>. 3.
Fig. 3.
Format is the same as in fig. 1. (A) Partial ML phylogeny of sxtG candidate proteins, the complete tree is in supplementary fig. S2, Supplementary Material online. (B) ML phylogeny of sxtB candidate proteins.

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