Bacillus thuringiensis Cry2Ab is active on Anopheles mosquitoes: single D block exchanges reveal critical residues involved in activity

FEMS Microbiol Lett. 2011 Dec;325(1):16-21. doi: 10.1111/j.1574-6968.2011.02403.x. Epub 2011 Oct 10.

Abstract

Cry2Aa exhibits dual activity to Lepidoptera and Diptera. Cry2Ab differs in amino acid sequence from Cry2Aa by 13% and has shown significant lepidopteran activity, but no mosquitocidal activity. Previous studies implicate 23 Cry2Aa specificity-conferring residues of domain II, which differ in Cry2Ab. Nine residues are putatively involved in conferring Cry2Aa dipteran specificity. To explore Cry2Ab dipteran toxicity, site-directed mutagenesis was employed to exchange Cry2Ab residues with Cry2Aa D (dipteran) block residues. Cry2Ab wild type demonstrated high toxicity (LC(50) of 540 ng mL(-1)) to Anopheles gambiae, but not to Aedes or Culex, within a 24-h time period. Cry2Ab should be reclassified as a dual active Cry toxin. Cry2Ab mutagenesis revealed critical residues for Cry2Ab protein function, as well as enhanced activity against the malarial mosquito, An. gambiae.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aedes / drug effects
  • Animals
  • Anopheles / drug effects*
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / genetics
  • Bacterial Proteins / pharmacology*
  • Culex / drug effects
  • Endotoxins / genetics
  • Endotoxins / pharmacology*
  • Hemolysin Proteins / genetics
  • Hemolysin Proteins / pharmacology*
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Mutant Proteins / genetics
  • Mutant Proteins / pharmacology
  • Protein Conformation
  • Structure-Activity Relationship

Substances

  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Endotoxins
  • Hemolysin Proteins
  • Mutant Proteins
  • insecticidal crystal protein, Bacillus Thuringiensis