Characterization of Echinostoma cinetorchis endoribonuclease, RNase H

Korean J Parasitol. 2017 Aug;55(4):451-455. doi: 10.3347/kjp.2017.55.4.451. Epub 2017 Aug 31.

Abstract

Echinostoma cinetorchis is an oriental intestinal fluke causing significant pathological damage to the small intestine. The aim of this study was to determine a full-length cDNA sequence of E. cinetorchis endoribonuclease (RNase H; EcRNH) and to elucidate its molecular biological characters. EcRNH consisted of 308 amino acids and showed low similarity to endoribonucleases of other parasites (<40%). EcRNH had an active site centered on a putative DDEED motif instead of DEDD conserved in other species. A recombinant EcRNH produced as a soluble form in Escherichia coli showed enzymatic activity to cleave the 3'-O-P bond of RNA in a DNA-RNA duplex, producing 3'-hydroxyl and 5'-phosphate. These findings may contribute to develop antisense oligonucleotides which could damage echinostomes and other flukes.

Keywords: Echinostoma cinetorchis; RNase H; endoribonuclease; localization; recombinant protein.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence / genetics*
  • DNA, Helminth / genetics*
  • Echinostoma / enzymology*
  • Oligonucleotides, Antisense
  • Ribonuclease H / chemistry
  • Ribonuclease H / genetics*
  • Sequence Analysis, DNA

Substances

  • DNA, Helminth
  • Oligonucleotides, Antisense
  • Ribonuclease H