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1.
Biophys J ; 99(2): 447-55, 2010 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-20643062

RESUMO

Bacteriophage T5 DNA ejection is a complex process that occurs on several timescales in vitro. By using a combination of bulk and single phage measurements, we quantitatively study the three steps of the ejection-binding to the host receptor, channel-opening, and DNA release. Each step is separately addressed and its kinetics parameters evaluated. We reconstruct the bulk kinetics from the distribution of single phage events by following individual DNA molecules with unprecedented time resolution. We show that, at the single phage level, the ejection kinetics of the DNA happens by rapid transient bursts that are not correlated to any genome sequence defects. We speculate that these transient pauses are due to local phase transitions of the DNA inside the capsid. We predict that such pauses should be seen for other phages with similar DNA packing ratios.


Assuntos
Bacteriófagos/metabolismo , DNA Viral/metabolismo , Modelos Biológicos , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Escherichia coli/metabolismo , Ativação do Canal Iônico , Cinética , Ligação Proteica , Fatores de Tempo
2.
J Phys Condens Matter ; 22(45): 454122, 2010 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-21339609

RESUMO

We experimentally study the statistical distributions and the voltage dependence of the unzipping time of 45 base-pair-long double-stranded DNA through a nanopore. We then propose a quantitative theoretical description considering the nanopore unzipping process as a random walk of the opening fork through the DNA sequence energy landscape biased by a time-fluctuating force. To achieve quantitative agreement fluctuations need to be correlated over the millisecond range and have an amplitude of order k(B)T/bp. Significantly slower or faster fluctuations are not appropriate, suggesting that the unzipping process is efficiently enhanced by noise in the kHz range. We further show that the unzipping time of short 15 base-pair hairpins does not always increase with the global stability of the double helix and we theoretically study the role of DNA elasticity on the conversion of the electrical bias into a mechanical unzipping force.


Assuntos
DNA/química , DNA/ultraestrutura , Modelos Químicos , Modelos Moleculares , Simulação por Computador , Campos Eletromagnéticos , Modelos Estatísticos , Conformação de Ácido Nucleico , Desnaturação de Ácido Nucleico , Porosidade , Estresse Mecânico
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