Bovine DNase I: Gene organization, mRNA expression, and changes in the topological distribution of the protein during apoptosis in lens epithelial cells

Alicia De María, Cristina Arruti

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Genomic DNA sequencing and alignment with the known DNase I mRNA showed that the bovine gene consists of 9 exons and that only the last 8 encode the protein, since initial ATG was found at exon II. RT-PCR was used to identify DNase I mRNA in lens epithelium in vivo and in cultured epithelial cells. We found DNase I transcripts having the same nucleotide sequence as the pancreas form and others lacking almost all exon V. The lens protein presented a slightly higher relative molecular weight than the pancreatic enzyme. Lens DNase I was located in secretory pathway organelles and excluded from the nucleus. Nevertheless, in apoptotic lens epithelial cells in vitro, DNase I translocated to the nucleus and co-localized with TUNEL positive chromatin aggregates. These results indicate that cells in the lens epithelium constitutively express DNase I, and suggest a direct involvement of this nuclease in the final phases of chromatin degradation.

Original languageEnglish
Pages (from-to)634-641
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume312
Issue number3
DOIs
StatePublished - Dec 19 2003

Keywords

  • Alternative splicing
  • Apoptosis
  • Chromatin condensation
  • DNase I
  • Endoplasmic reticulum
  • Lens epithelium
  • Nucleases
  • TUNEL

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