A Novel Intergenic ETnII-β Insertion Mutation Causes Multiple Malformations in Polypodia Mice

• DOI: 10.1371/journal.pgen.1003967
• Publication Date: 2013-12-05T21:30:13Z
• Authors: Jessica A. Lehoczky; Peedikayil E. Thomas; Kevin M. Patrie; Kailey M. Owens; Lisa M. Villarreal; Kenneth Galbraith; Joe Washburn; Craig N. Johnson; Bryant Gavino; Alexander D. Boro...; Kathleen J. Millen; Paul Wakenight; William Law; Margaret L. Van K...; Galina Gavrilina; Elizabeth D. Hughes; Thomas L. Saunders; Lesil Brihn; Joseph H. Nadeau; Jeffrey W. Innis
• Keywords: 0301 basic medicine; RNA Splicing; Chromosome Mapping; QH426-470; Mice; Mutagenesis, Insertional; 03 medical and health sciences; Phenotype; Calcium-Calmodulin-Dependent Protein Kinase Type 1; Gene Expression Regulation; Genes, X-Linked; Genetics; DNA Transposable Elements; Animals; Dual-Specificity Phosphatases; Research Article; Genes, Dominant

Mouse early transposon insertions are responsible for ~10% of spontaneous mutant phenotypes. We previously reported the phenotypes and genetic mapping of Polypodia, (Ppd), a spontaneous, X-linked dominant mutation with profound effects on body plan morphogenesis. Our new data shows that mutant mice are not born in expected Mendelian ratios secondary to loss after E9.5. In addition, we refined the Ppd genetic interval and discovered a novel ETnII-β early transposon insertion between the genes for Dusp9 and Pnck. The ETn inserted 1.6 kb downstream and antisense to Dusp9 and does not disrupt polyadenylation or splicing of either gene. Knock-in mice engineered to carry the ETn display Ppd characteristic ectopic caudal limb phenotypes, showing that the ETn insertion is the Ppd molecular lesion. Early transposons are actively expressed in the early blastocyst. To explore the consequences of the ETn on the genomic landscape at an early stage of development, we compared interval gene expression between wild-type and mutant ES cells. Mutant ES cell expression analysis revealed marked upregulation of Dusp9 mRNA and protein expression. Evaluation of the 5' LTR CpG methylation state in adult mice revealed no correlation with the occurrence or severity of Ppd phenotypes at birth. Thus, the broad range of phenotypes observed in this mutant is secondary to a novel intergenic ETn insertion whose effects include dysregulation of nearby interval gene expression at early stages of development.