Functional and phylogenetic analysis of a plant-inducible oligoribonuclease (orn) gene from an indigenous Pseudomonas plasmid

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Abstract
Application of a promoter-trapping strategy to identify plant-inducible genes carried on an indigenousPseudomonasplasmid, pQBR103, revealed the presence of a putative oligoribonuclease (orn) gene that encodes a highly conserved 3′ to 5′ exoribonuclease specific for small oligoribonucleotides. The deduced amino acid sequence of the plasmid-derivedorn(ornpl) showed three conserved motifs characteristic of Orn from both prokaryotes and eukaryotes. Deletion ofornplgenerated no observable phenotype, but inactivation of the chromosomal copy caused slow growth inPseudomonas putidaKT2440. This defect was fully restored by complementation withornfromEscherichia coli(ornE.coli). Plasmid-derivedornplwas capable of partially complementing theP. putida ornmutant, demonstrating functionality ofornpl. Phylogenetic analysis showed that plasmid-encoded Orn was distinct from Orn encoded by the chromosome of proteobacteria. A survey ofornplfrom relatedPseudomonasplasmids showed a sporadic distribution but no sequence diversity. These data suggest that theornplwas acquired by pQBR103 in a single gene-transfer event: the donor is unknown, but is unlikely to be a member of theProteobacteria.