A Novel Intergenic ETnII-β Insertion Mutation Causes Multiple Malformations in Polypodia Mice
| Autor: | William D. Law, Paul Wakenight, Alexander D. Borowsky, Margaret L. Van Keuren, Jessica A. Lehoczky, Joseph H. Nadeau, Elizabeth D. Hughes, Kailey M. Owens, Lisa M. Villarreal, Bryant J. Gavino, Galina B. Gavrilina, Jeffrey W. Innis, Peedikayil E. Thomas, Lesil E Brihn, Kenneth Galbraith, Craig N. Johnson, Thomas L. Saunders, Kathleen J. Millen, Joe Washburn, Kevin M. Patrie |
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| Rok vydání: | 2013 |
| Předmět: |
Cancer Research
lcsh:QH426-470 RNA Splicing Mutant Mutagenesis (molecular biology technique) Biology medicine.disease_cause Mice Genes X-Linked Gene expression Genetics medicine Animals Insertion Molecular Biology Gene Genetics (clinical) Ecology Evolution Behavior and Systematics Genes Dominant Regulation of gene expression Mutation Chromosome Mapping Molecular biology Mutagenesis Insertional lcsh:Genetics Phenotype Calcium-Calmodulin-Dependent Protein Kinase Type 1 Gene Expression Regulation DNA methylation DNA Transposable Elements Dual-Specificity Phosphatases Research Article |
| Zdroj: | PLoS Genetics, Vol 9, Iss 12, p e1003967 (2013) PLoS Genetics |
| ISSN: | 1553-7404 |
| Popis: | 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. Author Summary Mobile genetic elements, particularly early transposons (ETn), cause malformations by inserting within genes leading to disruption of exons, splicing or polyadenylation. Few mutagenic early transposon insertions have been found outside genes and the effects of such insertions on surrounding gene regulation is poorly understood. We discovered a novel intergenic ETnII-β insertion in the mouse mutant Polypodia (Ppd). We reproduced the mutant phenotype after engineering the mutation in wild-type cells with homologous recombination, proving that this early transposon insertion is Ppd. Mutant mice are not born in expected Mendelian ratios secondary to loss after E9.5. Embryonic stem cells from mutant mice show upregulated transcription of an adjacent gene, Dusp9. Thus, at an early and critical stage of development, dysregulated gene transcription is one consequence of the insertion mutation. DNA methylation of the ETn 5′ LTR is not correlated with phenotypic outcome in mutant mice. Polypodia is an example of an intergenic mobile element insertion in mice causing dramatic morphogenetic defects and fetal death. |
| Databáze: | OpenAIRE |
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