Structural and functional analysis of the symmetrical Type I restriction endonuclease R.EcoR124I_NT

ABSTRACT: Type I restriction-modification (RM) systems are comprised of two multi-subunit enzymes, the methyltransferase (∼160 kDa), responsible for methylation of DNA, and the restriction endonuclease (∼400 kDa), responsible for DNA cleavage. Both enzymes share a number of subunits. An engineered RM system, EcoR124I_NT, based on the N-terminal domain of the specificity subunit of EcoR124I was constructed that recognises the symmetrical sequence GAAN₇TTC and is active as a methyltransferase. Here, we investigate the restriction endonuclease activity of R.EcoR124I_NT in vitro and the subunit assembly of the multi-subunit enzyme. Finally, using small-angle neutron scattering and selective deuteration, we present a low-resolution structural model of the endonuclease and l

SUBMITTER: Taylor JE 

PROVIDER: S-EPMC3320862 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature