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Domain organization and metal ion requirement of the Type IIS restriction endonuclease MnlI
Author(s) -
Kriukiene Edita
Publication year - 2006
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2006.09.075
Subject(s) - nuclease , restriction enzyme , cleavage (geology) , dna , colicin , endonuclease , stereochemistry , biology , chemistry , biochemistry , microbiology and biotechnology , genetics , plasmid , paleontology , fracture (geology)
A two‐domain structure of the Type IIS restriction endonuclease MnlI has been identified by limited proteolysis. An N‐terminal domain of the enzyme mediates the sequence‐specific interaction with DNA, whereas a monomeric C‐terminal domain resembles bacterial colicin nucleases in its requirement for alkaline earth as well as transition metal ions for double‐ and single‐stranded DNA cleavage activities. The results indicate that the fusion of the non‐specific HNH‐type nuclease to the DNA binding domain had transformed MnlI into a Mg 2+ ‐, Ni 2+ ‐, Co 2+ ‐, Mn 2+ ‐, Zn 2+ ‐, Ca 2+ ‐dependent sequence‐specific enzyme. Nevertheless, MnlI retains a residual single‐stranded DNA cleavage activity controlled by its C‐terminal colicin‐like nuclease domain.