Abstract：
:Actinomycin D (ActD) is a small molecule with strong antibiotic and anticancer activity. However, its biologically relevant DNA-binding mechanism has never been resolved, with some studies suggesting that the primary binding mode is intercalation, and others suggesting that single-stranded DNA binding is most important. To resolve this controversy, we develop a method to quantify ActD's equilibrium and kinetic DNA-binding properties as a function of stretching force applied to a single DNA molecule. We find that destabilization of double stranded DNA (dsDNA) by force exponentially facilitates the extremely slow ActD-dsDNA on and off rates, with a much stronger effect on association, resulting in overall enhancement of equilibrium ActD binding. While we find the preferred ActD-DNA-binding mode to be to two DNA strands, major duplex deformations appear to be a pre-requisite for ActD binding. These results provide quantitative support for a model in which the biologically active mode of ActD binding is to pre-melted dsDNA, as found in transcription bubbles. DNA in transcriptionally hyperactive cancer cells will therefore likely efficiently and rapidly bind low ActD concentrations (≈ 10 nM), essentially locking ActD within dsDNA due to its slow dissociation, blocking RNA synthesis and leading to cell death.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Paramanathan T,Vladescu I,McCauley MJ,Rouzina I,Williams MCdoi
10.1093/nar/gks069subject
Has Abstractpub_date
2012-06-01 00:00:00pages
4925-32issue
11eissn
0305-1048issn
1362-4962pii
gks069journal_volume
40pub_type
杂志文章abstract：:Giant viruses have extraordinarily large dsDNA genomes, and exceptionally, they encode various components of the translation apparatus, including tRNAs, aminoacyl-tRNA synthetases and translation factors. Here, we focused on the elongation factor 1 (EF1) family of viral translational GTPases (trGTPases), using computa...
journal_title：Nucleic acids research
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abstract：:The role of Ribonuclease H1 (RNase H1) during primer removal and ligation at the mitochondrial origin of light-strand DNA synthesis (OriL) is a key, yet poorly understood, step in mitochondrial DNA maintenance. Here, we reconstitute the replication cycle of L-strand synthesis in vitro using recombinant mitochondrial p...
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doi：10.1093/nar/22.18.3706
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journal_title：Nucleic acids research
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pub_type： 杂志文章
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更新日期：2013-02-01 00:00:00
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journal_title：Nucleic acids research
pub_type： 杂志文章
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journal_title：Nucleic acids research
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journal_title：Nucleic acids research
pub_type： 杂志文章
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journal_title：Nucleic acids research
pub_type： 杂志文章
doi：10.1093/nar/4.5.1159-a
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journal_title：Nucleic acids research
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