Exploring the role of 5' alternative splicing and of the 3'-untranslated region of cathepsin B mRNA. Academic Article uri icon

Overview

abstract

  • The cysteine peptidase cathepsin B is responsible for connective tissue breakdown in several diseases. The pathological expression of cathepsin B may depend on the structure of its mRNA. We investigated the translational efficiency of the cathepsin B mRNA untranslated regions (UTRs) using fusion constructs to green fluorescent protein (GFP) and luciferase. Transfection of fusion constructs with GFP and luciferase containing the full-length 5'-UTR, the variant lacking exon 2, and that lacking exons 2 and 3 into mammalian cells, resulted in modulation of the biosynthetic rate of cathepsin B in a cell-specific manner. Constructs missing these exons were biosynthetically more efficient than the full-length counterpart. Luciferase was cloned upstream of the 3'-UTR, downstream of the 5'-UTR, or sandwiched between the 5'- and the 3'-UTR. The UTRs of cathepsin B downregulated luciferase biosynthesis moderately when present individually, with the 3'-UTR being more efficient than the 5'-UTR, and downregulated it even more when present simultaneously. A truncated cathepsin B-GFP chimeric product derived from the 5'-UTR missing exons 2 and 3 induced cell death. The increased biosynthetic rate and abnormal trafficking of cathepsin B observed in pathologies such as cancer and osteoarthritis may depend on alternative splicing of pre-mRNA.

publication date

  • July 1, 2003

Research

keywords

  • 3' Untranslated Regions
  • 5' Untranslated Regions
  • Alternative Splicing
  • Cathepsin B

Identity

Scopus Document Identifier

  • 0042856240

Digital Object Identifier (DOI)

  • 10.1515/BC.2003.113

PubMed ID

  • 12956417

Additional Document Info

volume

  • 384

issue

  • 7