ATP-type DNA ligase requires other proteins for its activity in vitro and its operon components for radiation resistance in Deinococcus radiodurans in vivo

Biochem Cell Biol. 2010 Oct;88(5):783-90. doi: 10.1139/o10-075.


A multiprotein DNA processing complex isolated from Deinococcus radiodurans contains the DNA repair protein PprA, an ATP-type DNA repair ligase (LigB) encoded by the drB0100 gene, and protein kinase activity. An ATP-dependent DNA end-joining activity was detected in the complex. To elucidate the function of the drB0100 gene, we generated the deletion mutant for the DR_B0100 ORF. The mutant exhibited a nearly 2-log cycle reduction in growth rate when exposed to a 10,000 Gray dose of γ-radiation, and a significant loss in mitomycin C and methylmethane sulphonate tolerance as compared with wild type. Functional complementation of these phenotypes required the wild-type copy of drB0100 along with other genes such as drb0099 and drb0098, organized downstream in the operon. The in vitro DNA ligase activity of LigB was stimulated severalfold by PprA in the presence of the recombinant DRB0098 protein. However, this activity did not improve when PprA was substituted with purified DRB0099 protein or when DRB0098 protein was substituted with the DRB0099 protein in the presence of PprA in solution. These results suggest that PprA and DRB0098 protein are required for LigB function. Furthermore, they also suggest that the LigB operon components contribute to radiation resistance and double-strand break (DSB) repair in D. radiodurans.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cell Survival / drug effects
  • Cell Survival / radiation effects
  • DNA Damage / drug effects
  • DNA Damage / genetics
  • DNA Damage / radiation effects
  • DNA Ligase ATP
  • DNA Ligases / genetics
  • DNA Ligases / metabolism*
  • DNA Repair / drug effects
  • DNA Repair / genetics
  • DNA Repair / radiation effects
  • DNA, Bacterial / genetics*
  • Deinococcus / enzymology
  • Deinococcus / radiation effects*
  • Gene Expression Regulation, Bacterial*
  • Mutation / genetics
  • Operon / genetics*
  • Radiation Tolerance*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism


  • Bacterial Proteins
  • DNA, Bacterial
  • Recombinant Proteins
  • DNA Ligases
  • DNA Ligase ATP