Inversion region for hypertension and brachydactyly on chromosome 12p features multiple splicing and noncoding RNA

Hypertension. 2008 Feb;51(2):426-31. doi: 10.1161/HYPERTENSIONAHA.107.101774. Epub 2007 Dec 17.

Abstract

Autosomal-dominant hypertension and brachydactyly (Online Mendelian Inheritance in Man 112410) is a prototype-translational research project. We used interphase fluorescent in situ hybridization and discovered complex rearrangements on chromosome 12p in 5 families but elucidated a common inverted region in the linkage interval. The inversion contains no known gene. However, we found 5 expressed sequence tags in databases. We used 5'- and 3'-Rapid Amplification of cDNA Ends PCR for elongation of the transcripts in phenotype-relevant tissue (fetal aorta, fetal brain, and fetal cartilage). We detected tissue-specific multiple splicing with different exon usage of 32 exons in the gene-related structure. These different transcripts lack both open reading frames and Kozak sequences. In vitro transcription/translation experiments did not identify any peptide-related molecules. We then performed quantitative RT-PCR to test for differential expression of the various spliced transcripts in the total fibroblast RNA of affected and nonaffected Turkish family members. Skin fibroblasts of affected individuals have a significantly increased proliferation rate compared with nonaffected individuals. Ten of 12 spliced exon combinations representing all of the spliced variants do not show a significantly different RNA expression rate. However, 2 RT-PCR products are exclusively expressed in nonaffected individuals. Both reverse transcription amplicons share 1 exon. This result is surprising because of the autosomal-dominant mode of inheritance of the trait. RNA secondary prediction of this single exon results in a stable stem-loop structure known to be essential for microRNA processing. We are pursuing the possibility of microRNA expression in affected patients that leads to complete down regulation of a spliced transcript.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing*
  • Cell Division
  • Chromosome Inversion*
  • Chromosome Mapping
  • Chromosome Painting
  • Chromosomes, Human, Pair 12*
  • Exons
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Gene Expression
  • Genes, Dominant
  • Humans
  • Hypertension / complications
  • Hypertension / genetics*
  • Interphase
  • Limb Deformities, Congenital / complications
  • Limb Deformities, Congenital / genetics*
  • Limb Deformities, Congenital / metabolism
  • Limb Deformities, Congenital / pathology
  • MicroRNAs / metabolism
  • RNA, Untranslated*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Skin / pathology

Substances

  • MicroRNAs
  • RNA, Untranslated