Domain organization and metal ion requirement of the Type IIS restriction endonuclease MnlI |
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Authors: | Kriukiene Edita |
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Affiliation: | Institute of Biotechnology, Graiciuno 8, Vilnius LT-02241, Lithuania. ediga@ibt.lt |
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Abstract: | 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. |
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Keywords: | Restriction endonuclease HNH motif Colicin Modular structure MnlI |
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