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Differential splicing and alternative polyadenylation generate multiple mimecan mRNA transcripts

E S Tasheva1, L M Corpuz, J L Funderburgh

  • 1Division of Biology, Kansas State University, Manhattan, Kansas 66506-4901, USA. est@ksu.edu

The Journal of Biological Chemistry
|January 24, 1998
PubMed
Summary

Researchers identified a novel splice variant of the mimecan (osteoglycin) gene, which is the predominant form in bovine cornea and sclera. Alternative polyadenylation also contributes to mimecan RNA diversity.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • The 25-kDa corneal keratan sulfate proteoglycan, known as mimecan, is a translation product of the osteoglycin gene.
  • Previous studies identified three mimecan RNA transcripts, but the mechanisms generating this diversity were unclear.

Purpose of the Study:

  • To investigate the mechanisms responsible for the generation of multiple mimecan RNA transcripts.
  • To characterize splice variants and alternative polyadenylation in mimecan gene expression.

Main Methods:

  • Ribonuclease protection analysis (RPA) and reverse transcription-polymerase chain reaction (RT-PCR) were used to analyze bovine corneal mRNA.
  • cDNA cloning was employed to identify novel sequences in the mimecan gene.

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Main Results:

  • A predominant mimecan mRNA splice variant lacking 278 base pairs in the 5'-untranslated region was detected in bovine cornea and sclera.
  • Two additional cDNA clones revealed novel sequences at the 3'-end of mimecan cDNA.
  • Alternative selection of three canonical polyadenylation sites in the 3'-untranslated region was confirmed.

Conclusions:

  • The diversity of mimecan RNA isoforms arises from both alternative splicing and alternative polyadenylation.
  • These findings highlight a previously unrecognized level of complexity in mimecan gene expression within the same tissue.