Analysis of retrotransposon families in genomic DNA by two-dimensional restriction mapping: Detection of VL30 insertions in mouse thymic lymphoma

Abstract

Reinsertion of mammalian retrotransposable genetic elements is known to be causally associated with tumourigenesis, typically through mechanisms involving insertional deregulation of cellular protooncogene expression. We report here on the application of a two-dimensional restriction mapping-Southern hybridisation approach for analysis of retrotransposon families of low to moderate genetic complexity, which is particularly suited to pairwise comparisons between DNA samples. By using this method, non-constitutional mink-cell-focus-forming type retro-elements were readily detectable in AKR mouse thymic lymphomas against a background of approx. 30 related elements in control DNA. However, in the WEHI 3B myeloid leukaemia cell line, the resolution of two-dimensional mapping permitted detection of only occasional reinsertions of intracisternal A particle retro-elements (genetic complexity: 10 3). In analysing the VL30 family of retrotransposon (genetic complexity: 150) we developed a strategy for identifying the known transcriptionally active sub-set of these elements in genomic DNA through the generation of an internal, diagnostic restriction fragment. Moreover, in some cases of thymic lymphoma, several candidate re-insertions of VL30 elements were detected, consistent with a suggested role for retrotransposition of this class of element in lymphomagenesis of retroviral aetiology.