The Relaxase of the Rhizobium Etli Symbiotic Plasmid Shows Nic Site Cis-acting Preference

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 2006 Aug 19

PMID 16916896

Citations 12

Authors

Daniel Perez-Mendoza

Maria Lucas

Socorro Munoz

Jose A Herrera-Cervera

Jose Olivares

Fernando de la Cruz

Juan Sanjuan

Affiliations

Soon will be listed here.

Abstract

Genetic and biochemical characterization of TraA, the relaxase of symbiotic plasmid pRetCFN42d from Rhizobium etli, is described. After purifying the relaxase domain (N265TraA), we demonstrated nic binding and cleavage activity in vitro and thus characterized for the first time the nick site (nic) of a plasmid in the family Rhizobiaceae. We studied the range of N265TraA relaxase specificity in vitro by testing different oligonucleotides in binding and nicking assays. In addition, the ability of pRetCFN42d to mobilize different Rhizobiaceae plasmid origins of transfer (oriT) was examined. Data obtained with these approaches allowed us to establish functional and phylogenetic relationships between different plasmids of this family. Our results suggest novel characteristics of the R. etli pSym relaxase for previously described conjugative systems, with emphasis on the oriT cis-acting preference of this enzyme and its possible biological relevance.

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