Identification and characterization of a new Leishmania major specific 3′nucleotidase/nuclease protein

We report the characterization of a new Leishmania major gene, lmaj3′nt/nu, encoding a 382 amino acids protein, Lmaj3′NT/NU, that belongs to the 3′nucleotidase/nuclease family. Interestingly, sequence and phylogenetic analysis show that this protein is Leishmania major specific and thus constitutes a new 3′nucleotidase/nuclease subgroup. Lmaj3′NT/NU displays nuclease enzymatic activity and Western blot analysis shows that it is exclusively expressed in promastigotes. Immunofluorescence microscopy using a specific anti-Lmaj3′NT/NU shows that the protein has a plasma membrane localization. Surprisingly, contrary to the previously described Leishmania mexicana 3′NT/NU, lmaj3′nt/nu is not up-regulated when parasites are cultured under purine starvation conditions. Together, these findings suggest Lmaj3′NT/NU may constitute a new important compound of the L. major purine scavenging pathway and could be involved in sandfly parasite survival and colonization.