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Characterization of restriction-modification enzymes Cfr13 I from Citrobacter freundii RFL13

J B Bitinaité, S J Klimasauskas, V V Butkus

    FEBS Letters
    |March 25, 1985
    PubMed

    Insights

    The RCfr13 I restriction enzyme and MCfr13 I methylase from Citrobacter freundii RFL13 were characterized. RCfr13 I exhibits sensitivity to specific DNA methylation patterns, including those from E. coli dcm methylase.

    Area of Science:

    • Molecular Biology
    • Enzymology
    • Genetics

    Background:

    • Restriction-modification systems are crucial for bacterial defense and genome regulation.
    • Understanding enzyme specificity is key to molecular biology applications.

    Purpose of the Study:

    • To characterize the RCfr13 I restriction enzyme and MCfr13 I methylase from Citrobacter freundii RFL13.
    • To investigate the substrate specificity of RCfr13 I, particularly its sensitivity to DNA methylation.

    Main Methods:

    • Isolation and characterization of RCfr13 I restriction enzyme and MCfr13 I methylase.
    • Enzymatic assays to determine recognition sequences and cleavage sites.
    • Analysis of enzyme activity on methylated and hemimethylated DNA substrates.

    Main Results:

    • RCfr13 I recognizes and cleaves the 5'-G GNCC sequence.
    • MCfr13 I methylase modifies the internal cytosine to 5'-GGNm5CC.
    • RCfr13 I is sensitive to substrate modification by MCfr13 I, MCfr10 I, and MHpa II.

    Conclusions:

    • RCfr13 I displays significant sensitivity to specific DNA methylation patterns, including hemimethylation.
    • The findings suggest RCfr13 I is sensitive to methylation by E. coli dcm methylase in overlapping sites.

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