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Background   PIF/Harbinger  is the most recently discovered DNA transposon superfamily and is now known to populate genomes from fungi to plants to animals. Mobilization of superfamily members requires two separate element-encoded proteins (ORF1 and TPase). Members of this superfamily also mobilize  Tourist -like miniature inverted repeat transposable elements (MITEs), which are the most abundant transposable elements associated with the genes of plants, especially the cereal grasses. The phylogenetic analysis of many plant genomes indicates that MITEs can amplify rapidly from one or a few elements to hundreds or thousands.  The most active DNA transposon identified to date in plants or animals is  mPing , a rice  Tourist -like MITE that is a deletion derivative of the autonomous  Ping  element.  Ping  and the closely related  Pong  are the only known naturally active  PIF/Harbinger  elements. Some rice strains accumulate ~40 new  mPing  insertions per plant per generation. In this study we report the development of a yeast transposition assay as a first step in deciphering the mechanism underlying the amplification of  Tourist -MITEs.    Results  The ORF1 and TPase proteins encoded by  Ping  and  Pong  have been shown to mobilize  mPing  in rice and in transgenic  Arabidopsis . Initial tests of the native proteins in a yeast assay resulted in very low transposition. Significantly higher activities were obtained by mutation of a putative nuclear export signal (NES) in the TPase that increased the amount of TPase in the nucleus. When introduced into  Arabidopsis , the NES mutant protein also catalyzed higher frequencies of  mPing  excision from the  gfp  reporter gene. Our yeast assay retains key features of excision and insertion of  mPing  including precise excision, extended insertion sequence preference, and a requirement for two proteins that can come from either  Ping  or  Pong  or both elements.    Conclusions  The yeast transposition assay provides a robust platform for analysis of the mechanism underlying transposition catalyzed by the two proteins of  PIF/Harbinger  elements. It recapitulates all of the features of excision and reinsertion of  mPing  as seen in plant systems. Furthermore, a mutation of a putative NES in the TPase increased transposition both in yeast and plants.

Transposition of the Tourist-MITE mPing in yeast: an assay that retains key features of catalysis by the class 2 PIF/Harbinger superfamily