Evilevitch Alex, Fang Li Tai, Yoffe Aron M, Castelnovo Martin, Rau Donald C, Parsegian V Adrian, Gelbart William M, Knobler Charles M
Department of Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, Lund, Sweden.
Biophys J. 2008 Feb 1;94(3):1110-20. doi: 10.1529/biophysj.107.115345. Epub 2007 Sep 21.
Recent work has shown that pressures inside dsDNA phage capsids can be as high as many tens of atmospheres; it is this pressure that is responsible for initiation of the delivery of phage genomes to host cells. The forces driving ejection of the genome have been shown to decrease monotonically as ejection proceeds, and hence to be strongly dependent on the genome length. Here we investigate the effects of ambient salts on the pressures inside phage-lambda, for the cases of mono-, di-, and tetravalent cations, and measure how the extent of ejection against a fixed osmotic pressure (mimicking the bacterial cytoplasm) varies with cation concentration. We find, for example, that the ejection fraction is halved in 30 mM Mg(2+) and is decreased by a factor of 10 upon addition of 1 mM spermine. These effects are calculated from a simple model of genome packaging, using DNA-DNA repulsion energies as determined independently from x-ray diffraction measurements on bulk DNA solutions. By comparing the measured ejection fractions with values implied from the bulk DNA solution data, we predict that the bending energy makes the d-spacings inside the capsid larger than those for bulk DNA at the same osmotic pressure.
近期研究表明,双链DNA噬菌体衣壳内部的压力可高达数十个大气压;正是这种压力导致噬菌体基因组向宿主细胞的传递起始。研究表明,随着基因组释放过程的推进,驱动基因组释放的力会单调下降,因此强烈依赖于基因组长度。在此,我们研究了单价、二价和四价阳离子情况下,环境盐对λ噬菌体内部压力的影响,并测量了在固定渗透压(模拟细菌细胞质)下,基因组释放程度随阳离子浓度的变化情况。例如,我们发现,在30 mM Mg(2+)中,释放分数减半,而加入1 mM精胺后,释放分数降低了10倍。这些效应是通过一个简单的基因组包装模型计算得出的,该模型使用了从大量DNA溶液的X射线衍射测量中独立确定的DNA-DNA排斥能。通过将测量的释放分数与大量DNA溶液数据所暗示的值进行比较,我们预测,在相同渗透压下,弯曲能会使衣壳内部的d间距大于大量DNA的d间距。