Protection of particular cleavage sites of restriction endonucleases by distamycin A and actinomycin D

Abstract

It is shown here that distamycin A and actinomycin D can protect the recognition sites of endo R.EcoRI, EcoRII, HindII, HindIII, HpaI and HpaII from the attack of these restriction endonucleases. At proper distamycin concentrations only two endo R.EcoRI sites of phage lambda DNA are available for the restriction enzyme--sRI1 and sRI4. This phenomenon results in the appearance of larger DNA fragments comprising several consecutive fragments of endo R.EcoRI complete cleavage. The distamycin fragments isolated from the agarose gels can be subsequently cleaved by endo R.EcoRI with the yield of the fragments of complete digestion. We have compared the effect of distamycin A and actinomycin D on a number of restriction endonucleases having different nucleotide sequences in the recognition sites and established that antibiotic action depends on the nucleotide sequences of the recognition sites and their closest environment

The use of antibiotics actinomycin D and distamycin A for mapping of phage lambda HindIII fragments

“…This allows us to obtain a set of larger overlapping fragments of DNA (hereafter we shall term them as distamycin and actinomycin protected fragments) and to establish the location of all Hind111 fragments on the physical map of phage lambda DNA. The data reported here and earlier [3,4] lead to a conclusion that the anti- …”

“…This interaction is rather selective: distamycin forms complexes preferentially with d(A/T) rich regions [ 11 whereas actinomycin binds d(G/C) rich regions [2] of DNA. This property has offered a possibility to use these antibiotics for effective and highly selective protection of the cleavage sites recognized by restriction endonucleases [3,4]. The recognition site of endo R. HindIII [5] 5' AAGCTT 3'TTCGAA may be protected by both actinomycin D and distamycin A as long as it contains TT and GC sequences favourable for interaction of distamycin and actinomycin with template DNA.…”

“…The recognition site of endo R. HindIII [5] 5' AAGCTT 3'TTCGAA may be protected by both actinomycin D and distamycin A as long as it contains TT and GC sequences favourable for interaction of distamycin and actinomycin with template DNA. It has been demonstrated [4] that different EcoRI recognition sites on lambda DNA may be effectively blocked by different antibiotic concentrations due to the peculiarities of the environment of different recognition sites. In the present paper it is shown that certain endo R. Hind111 sites of phage lambda DNA may be effectively protected with actinomycin whereas some other sites are protected with distamycin.…”

Distamycin-induced inhibition of homeodomain-DNA complexes.

“…It is known that rates of enzymatic cleavage of DNA can vary widely (Nosikov et al, 1976;Kuroyedov et al, 1977;Malcolm and Moffatt, 1981), e.g. the difference between the most and the least susceptible EcoRI sites in bacteriophage X DNA is -10-fold (Thomas and Davis, 1975).…”

Structure at restriction endonuclease MboI cleavage sites protected by actinomycin D or distamycin A

“…Nosikov et al reported [2] that the DNA binding compounds actinomycin D and distamycin A inhibit cleavage of DNA by a number of different restriction endonucleases. Actinomycin D intercalates between the bases of dG-dC dinucleotide residues in duplex DNA [3], whereas distamycin A binds by both electrostatic and non-ionic forces to runs of dT residues [4,5].…”