Inhibition of DNA replication by berenil in plasmids containing poly(dA)poly(dT) sequences

Lorenzo Coates, Emeka V. Ikpeazu, Yeeru Chen, Manuel S. Valenzuela

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Abstract

The effect of berenil on plasmid DNA replication was studied on pBR322-derived plasmids containing poly(dA)poly(dT) sequences. In comparison to the parental plasmid pBR322, plasmid pKH47 harboring 100 bp of poly(dA)poly(dT) at the PvuII site showed a decrease in plasmid yield in the presence of berenil. This effect was also observed in pVL26, a related plasmid in which the location of the poly(dA)poly(dT) region had been shifted to the EcoRV site in pBR322. [3H]Thymidine incorporation experiments indicated that DNA synthesis may be affected in these plasmids in the presence of the drug. Bromodeoxyuridine incorporation experiments coupled to Cs2SO4 equilibrium density gradient centrifugation indicated that the lower plasmid yield was due to an inhibition of DNA replication by berenil. We have also found that berenil induces DNA degradation in plasmids containing the homopolymer. Our studies strongly suggest that the effect of berenil on plasmid replication and DNA stability results from its binding to the poly(dA)poly(dT) region present in these plasmids. Moreover, we have found a correlation between the position of the poly(dA)poly(dT) region and this inhibitory effect. Thus, plasmid pKH47, containing the poly(dA)poly(dT) region most proximal to the origin of pBR322 replication, was most severely affected.

Original languageEnglish (US)
Pages (from-to)120-128
Number of pages9
JournalPlasmid
Volume47
Issue number2
DOIs
StatePublished - Jan 1 2002
Externally publishedYes

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Keywords

  • A-T-rich sequences
  • Berenil
  • Plasmid DNA replication

ASJC Scopus subject areas

  • Molecular Biology

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