In silico prediction of active site and in vitro DNase and RNase activities of Helicoverpa-inducible pathogenesis related-4 protein from Cicer arietinum

Int J Biol Macromol. 2018 Jul 1:113:869-880. doi: 10.1016/j.ijbiomac.2018.03.027. Epub 2018 Mar 7.

Abstract

Plants are endowed with an innate immune system, which enables them to protect themselves from pest and pathogen. The participation of pathogenesis-related (PR) proteins is one of the most crucial events of inducible plant defense response. Herein, we report the characterization of CaHaPR-4, a Helicoverpa-inducible class II PR-4 protein from chickpea. Bioinformatic analysis of CaHaPR-4 protein indicated the presence of a signal peptide, barwin domain but it lacks the chitin-binding site/hevein domain. The recombinant CaHaPR-4 is bestowed with RNase and bivalent ion-dependent DNase activity. Further, the RNA and DNA binding sites were identified and confirmed by analyzing interactions between mutated CaHaPR-4 with the altered active site and ribonuclease inhibitor, 5'ADP and DNase inhibitor, 2‑nitro‑5‑thiocyanobenzoic acid (NTCB) using 3D modeling and docking studies. Moreover, CaHaPR-4 shows antifungal activity as well as growth inhibiting properties against neonatal podborer larvae. To the best of our knowledge, this is the first report of a PR-4 showing RNase, DNase, antifungal and most importantly insect growth inhibiting properties against Helicoverpa armigera simultaneously.

Keywords: Antifungal; Barwin domain; Chickpea; DNase; Helicoverpa; Insect growth inhibition; PR-4; RNase.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Catalytic Domain*
  • Cicer / enzymology*
  • Computer Simulation*
  • Deoxyribonucleases / antagonists & inhibitors
  • Deoxyribonucleases / metabolism*
  • Enzyme Inhibitors / metabolism
  • Enzyme Inhibitors / pharmacology
  • Fusarium / drug effects
  • Lepidoptera / drug effects
  • Lepidoptera / growth & development
  • Models, Molecular
  • Mutation
  • Phylogeny
  • Plant Proteins / chemistry*
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Plant Proteins / pharmacology
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Ribonucleases / metabolism*
  • Sequence Analysis
  • Thiocyanates / metabolism

Substances

  • Enzyme Inhibitors
  • Plant Proteins
  • Thiocyanates
  • Deoxyribonucleases
  • Ribonucleases
  • 2-nitro-5-thiocyanobenzoic acid