Mapping DNA cleavage by the Type ISP restriction-modification enzymes following long-range communication between DNA sites in different orientations.
Bottom Line: By following communication between sites in both head-to-head and head-to-tail orientations, we could show that motor activity leads to activation of the nuclease domains via distant interactions of the helicase or MTase-TRD.Direct nuclease dimerization is not required.To help explain the observed cleavage patterns, we also used exonuclease footprinting to demonstrate that individual Type ISP domains can swing off the DNA.
Affiliation: DNA-Protein Interactions Unit, School of Biochemistry, University of Bristol, Bristol BS8 1TD, UK.Show MeSH
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Mentions: The 157 bp linear one-site LlaGI DNA (Figure 7) was generated from the 1785–1941 region of pUC19 using primers FOli LlaGI +75 (5′-TCCAGATTTATCAGCAATAAACCAGCCAGCC-3′) and ROli LlaGI -75 (5′-AGCAATGGCAACAACGTTGCGCAAACTATTAAC-3′).
Affiliation: DNA-Protein Interactions Unit, School of Biochemistry, University of Bristol, Bristol BS8 1TD, UK.