Protein NCRII-18: the role of gene fusion in the molecular evolution of restriction endonucleases

FEBS Lett. 2017 Jun;591(12):1702-1711. doi: 10.1002/1873-3468.12669. Epub 2017 Jun 14.

Abstract

This work first constructed the fusion protein NCRII-18 by fusing the restriction endonuclease Ecl18kI gene and part of the gene coding for the N-terminal domain of the endonuclease EcoRII. The fusion of the EcoRII N-terminal domain leads to a change in the properties of the recombinant protein. Unlike Ecl18kI, which made the basis of NCRII-18, the fusion protein predominantly recognizes the CCWGG sites, having lost the capability of interacting with the CCSGG sites. Experimental data support the hypothesis of a close evolutionary relationship between type IIE and IIP restriction endonucleases via a recombination between domains with active site structure and elements for recognition with domains responsible for recognition of DNA sequences.

Keywords: DNA-protein interactions; domain organization; enterobacteria; fusion protein; restriction endonucleases.

MeSH terms

  • Artificial Gene Fusion
  • Binding Sites
  • Catalytic Domain
  • DNA Restriction Enzymes / chemistry
  • DNA Restriction Enzymes / genetics
  • DNA Restriction Enzymes / metabolism
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / metabolism
  • Deoxyribonucleases, Type II Site-Specific / chemistry
  • Deoxyribonucleases, Type II Site-Specific / genetics*
  • Deoxyribonucleases, Type II Site-Specific / metabolism
  • Electrophoretic Mobility Shift Assay
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics*
  • Escherichia coli Proteins / metabolism
  • Evolution, Molecular*
  • Gene Fusion*
  • Models, Genetic*
  • Nucleotide Motifs
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Protein Conformation
  • Protein Engineering
  • Recombinant Fusion Proteins / metabolism
  • Recombination, Genetic
  • Substrate Specificity

Substances

  • DNA, Bacterial
  • Escherichia coli Proteins
  • Peptide Fragments
  • Recombinant Fusion Proteins
  • DNA Restriction Enzymes
  • endodeoxyribonuclease Ecl18kI
  • CCWGG-specific type II deoxyribonucleases
  • Deoxyribonucleases, Type II Site-Specific