The splicing and fate of ADAM33 transcripts in primary human airways fibroblasts

Am J Respir Cell Mol Biol. 2004 Jul;31(1):13-21. doi: 10.1165/rcmb.2003-0330OC. Epub 2004 Jan 23.

Abstract

The ADAM (A Disintegrin and Metalloprotease) family of Zn++-dependent metalloproteases are multidomain proteins involved in diverse cellular activities. Polymorphic variation in ADAM33 is strongly associated with asthma and bronchial hyperresponsiveness. Identification of those isoforms of ADAM33 that are expressed in airways is fundamental to dissecting the role of ADAM33 in asthma. Analysis of primary human airways fibroblasts has shown the presence of a number of alternatively spliced forms of ADAM33, including one encoding a putative secreted variant, and many transcripts lacking the metalloproteinase domain. The relative abundance of these transcripts has been quantified using reverse transcription real-time polymerase chain reaction, in both nuclear and cytoplasmic fractions of RNA. These results demonstrate that a number of splice variants of ADAM33 are transported into the cytoplasm. Ninety percent of ADAM33 mRNA is retained in the nucleus and the subtle differences in the composition of nuclear and cytoplasmic RNA suggest important events in the splicing and selection of ADAM33 transcripts. Western blot analysis confirmed that several protein isoforms of ADAM33 are expressed in primary airways fibroblasts. These findings demonstrate that ADAM33 exists in multiple isoforms, suggesting that it is a complex molecule that plays multiple roles within mesenchymal cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins
  • Adult
  • Alternative Splicing / genetics
  • Asthma / genetics*
  • Asthma / metabolism
  • Asthma / physiopathology
  • Bronchi / cytology
  • Bronchi / metabolism*
  • Bronchi / physiopathology
  • Cell Nucleus / genetics
  • Cells, Cultured
  • Cytoplasm / genetics
  • Exons / genetics
  • Fibroblasts / cytology
  • Fibroblasts / metabolism*
  • Gene Expression Regulation / genetics
  • Humans
  • Metalloendopeptidases / biosynthesis
  • Metalloendopeptidases / genetics*
  • Polymorphism, Genetic / genetics
  • Protein Isoforms / genetics
  • Protein Structure, Tertiary / genetics
  • RNA, Messenger / metabolism*
  • Transcription, Genetic / genetics

Substances

  • Protein Isoforms
  • RNA, Messenger
  • ADAM Proteins
  • ADAM33 protein, human
  • Metalloendopeptidases