Molecular cloning and RNA expression of a novel Drosophila calpain, Calpain C

Cesare Spadoni, Attila Farkas, Rita Sinka, Peter Tompa, Peter Friedrich

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The calpains are Ca2+-activated cysteine proteases whose biochemical properties have been extensively characterized in vitro. Less is known, however, about the physiological role of calpains. In this respect, Drosophila melanogaster is a useful experimental organism to study calpain activity and regulation in vivo. The sequencing of the fly genome has been recently completed and a novel calpain homologue has been identified in the CG3692 gene product. We embarked on the cloning and characterization of this putative novel calpain. We demonstrate that the actual calpain is different from the predicted protein and we provide experimental evidence for the correction of the genomic annotation. This novel protein, Calpain C, must be catalytically inactive, having mutated active site residues but is otherwise structurally similar to the other known fly calpains. Moreover, we analysed Calpain C RNA expression during Drosophila development by RT-PCR and RNA in situ hybridization, which revealed strong expression in the salivary glands.

Original languageEnglish
Pages (from-to)343-349
Number of pages7
JournalBiochemical and biophysical research communications
Volume303
Issue number1
DOIs
Publication statusPublished - Mar 28 2003

Keywords

  • Calcium
  • Calpain
  • Cloning
  • Drosophila
  • Genomic annotation
  • In situ hybridization
  • RNA
  • RT-PCR

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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