Recombination and selectional forces in cyanopeptolin NRPS operons from highly similar, but geographically remote Planktothrix strains.
BACKGROUND
Cyanopeptolins are nonribosomally produced heptapetides showing a highly variable composition. The cyanopeptolin synthetase operon has previously been investigated in three strains from the genera Microcystis, Planktothrix and Anabaena. Cyanopeptolins are displaying protease inhibitor activity, but the biological function(s) is (are) unknown. Cyanopeptolin gene cluster variability and biological functions of the peptide variants are likely to be interconnected.


RESULTS
We have investigated two cyanopeptolin gene clusters from highly similar, but geographically remote strains of the same genus. Sequencing of a nonribosomal peptide synthetase (NRPS) cyanopeptolin gene cluster from the Japanese strain Planktothrix NIES 205 (205-oci), showed the 30 kb gene cluster to be highly similar to the oci gene cluster previously described in Planktothrix NIVA CYA 116, isolated in Norway. Both operons contained seven NRPS modules, a sulfotransferase (S) and a glyceric acid loading (GA)-domain. Sequence analyses showed a high degree of conservation, except for the presence of an epimerase domain in NIES 205 and the regions around the epimerase, showing high substitution rates and Ka/Ks values above 1. The two strains produce almost identical cyanopeptolins, cyanopeptolin-1138 and oscillapeptin E respectively, but with slight differences regarding the production of minor cyanopeptolin variants. These variants may be the result of relaxed adenylation (A)-domain specificity in the nonribosomal enzyme complex. Other genetic markers (16S rRNA, ntcA and the phycocyanin cpcBA spacer) were identical, supporting that these geographically separated Planktothrix strains are closely related.


CONCLUSIONS
A horizontal gene transfer event resulting in exchange of a whole module-encoding region was observed. Nucleotide statistics indicate that both purifying selection and positive selection forces are operating on the gene cluster. The positive selection forces are acting within and around the epimerase insertion while purifying selection conserves the remaining (major) part of the gene cluster. The presence of an epimerase in the gene cluster is in line with the D-configuration of Htyr, determined experimentally in oscillapeptin E in a previous study.
Rounge
Trine B
TB
University of Oslo, Department of Biology, Centre for Ecological and Evolutionary Synthesis, 0316 Oslo, Norway.t.b.rounge@bio.uio.no
Rohrlack
Thomas
T
Kristensen
Tom
T
Jakobsen
Kjetill S
KS
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D017124:Conserved Sequence; D000458:Cyanobacteria / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D019143:Evolution, Molecular; D005798:Genes, Bacterial; D014644:Genetic Variation; D005810:Multigene Family; D009876:Operon; D010453:Peptide Synthases / Q000235:genetics*; D010456:Peptides, Cyclic / Q000235:genetics*; D010802:Phylogeny; D011995:Recombination, Genetic; D012641:Selection, Genetic; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D010456:Peptides, Cyclic; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase