Nucleic Acids Research, 1995, Vol. 23, No. 10 1657-1663
© 1995
MOLECULAR BIOLOGY |
Distamycin A and tallimustine inhibit TBP binding and basal in vitro transcription
Dipartimento di Genetica e Biologia dei Microrganismi Università di Milano, Via Celoria 26, 20133 Milano, Italy 1Institut de Genetique et de Biologie Moleculaire et Cellulaire 1 Rue L.Fries, BP 163, 67404 Illkirch CU Strasbourg, France 2lstituto di Ricerche Farmacologiche ‘Mario Negri’ Via Eritrea 62, 20157 Milano, Italy 3Pharmacia/Farmitalia Carlo Erba, Nerviano Research Center Via Papa Giovanni XXIII 68, 20014 Nerviano, Italy
* To whom correspondence should be addressed
Received March 2, 1995. Revised April 5, 1995. Accepted April 5, 1995.
The antibiotic distamycin A is a DNA minor groove binding drug (MGB) that recognizes a stretch of a least four ATs. The alkylating benzoyl mustard derivative tallimustine (FCE 24517) has powerfull anti-tumor activity. Using the electrophoretic mobility shift assay (EMSA) we determined that both compounds can prevent binding of TBP and, with 10-fold higher concentration, TBP-TFIIA (DA) and TBP-TRIA-TFIIB (DAB) to a TATA box. Once formed, the DA and DAB complexes are more resistant to MGB challenge. Both drugs can inhibit basal In vitro transcription of a minimal TATA-containlng promoter and similar concentrations are necessary for binding and transcrip-tional inhibition. Tallimustine shows strong selectivity by decreasing only correctly initiated transcripts. Even at high doses (20µM), however, they cannot disturb a competent pre-initiation complex or Pol II progression. This functional in vitro model will provide a way to investigate the activity of sequence-specific DNA binding drugs with potential anti-viral and anti-tumour activity and to develop novel more selective compounds.
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