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Nucleic Acids Research, 1981, Vol. 9, No. 15 3695-3705
© 1981

ENZYMOLOGY

Specific and reversible inhibition of the blunt end joining activity of the T4 DNA ligase

Luca Ferretti and Vittorio Sgaramella

Istituto di Genetica Biochimica ed Evoluzionistica C.N.R., Via S.Epifanio 14, 27100 Pavia, Italy

Received May 5, 1981. Specific, complete and reversible inhibition of the joining of blunt ended DNA duplexes catalyzed by the T4 DNA ligase can be obtained by using ATP, the enzyme cofactor, at concentrations of 5 mM and higher.

On cohesive DNA ends, 5 mM ATF, which completely inhibits blunt end ligation, brings about only a limited (8%) reduction in the level of joining obtainable under optimal ATP concentration (O,5 mM or lower). A similar but less drastic uncoupling of the two kinds of joining can be achieved at lower ATP concentration (2,5 mM) using 1 mM Mg++

The joining of DNA blunt ends can also be inhibited almostcompletely by 10 mM spermidine, without noticeable effect on thejoining of cohesive termini.


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