Abstract:
:Endonucleases that generate double-strand breaks in DNA often possess two identical subunits related by rotational symmetry, arranged so that the active sites from each subunit act on opposite DNA strands. In contrast to many endonucleases, Type IIP restriction enzyme BcnI, which recognizes the pseudopalindromic sequence 5'-CCSGG-3' (where S stands for C or G) and cuts both DNA strands after the second C, is a monomer and possesses a single catalytic center. We show here that to generate a double-strand break BcnI nicks one DNA strand, switches its orientation on DNA to match the polarity of the second strand and then cuts the phosphodiester bond on the second DNA strand. Surprisingly, we find that an enzyme flip required for the second DNA strand cleavage occurs without an excursion into bulk solution, as the same BcnI molecule acts processively on both DNA strands. We provide evidence that after cleavage of the first DNA strand, BcnI remains associated with the nicked intermediate and relocates to the opposite strand by a short range diffusive hopping on DNA.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Sasnauskas G,Kostiuk G,Tamulaitis G,Siksnys Vdoi
10.1093/nar/gkr588subject
Has Abstractpub_date
2011-11-01 00:00:00pages
8844-56issue
20eissn
0305-1048issn
1362-4962pii
gkr588journal_volume
39pub_type
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