Identification and characterisation of a RAD51 gene from Leishmania major.

The RAD51 gene is a homologue of Escherichia coli recA which plays a central role in homologous recombination and DNA repair. This paper describes the identification of the RAD51 gene from the trypanosomatid parasite Leishmania major. The LmRAD51 gene codes for a 377 amino acid polypeptide with a predicted molecular mass of 41259 Da that is highly homologous to the Rad51 family of proteins. Recombinant L. major Rad51 protein (LmRad51) was over-expressed in a bacterial expression system, purified to homogeneity and shown to bind DNA and exhibit DNA-stimulated ATPase activity, consistent with previously reported biochemical characteristics of Rad51 protein. Although LmRad51 expression is below the level of detection in exponentially growing cultures of Leishmania, high levels of LmRad51 mRNA and protein expression can be detected following exposure to the DNA-damaging agent phleomycin. LmRAD51 is one of the first examples of a DNA damage-inducible gene to be characterised in Leishmania, and will be invaluable in studying the contribution of homologous recombination to Leishmania virulence.