Sun M, Serwer P
Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760, USA.
Biophys J. 1997 Feb;72(2 Pt 1):958-63. doi: 10.1016/s0006-3495(97)78730-7.
When packaged in a bacteriophage capsid, double-stranded DNA occupies a cavity whose volume is roughly twice the volume of the DNA double helix. The data thus far have not revealed whether the compactness of packaged bacteriophage DNA is achieved by folding of the DNA, undirectional winding of the DNA, or a combination of both folding and winding. To assist in discriminating among these possibilities, the present study uses electron microscopy, together with ultraviolet light-induced DNA-DNA cross-linking, to obtain the following information about the conformation of DNA packaged in the comparatively large bacteriophage, G: 1) At the periphery of some negatively stained particles of bacteriophage G, electron microscopy reveals standards of DNA that are both parallel to each other and parallel to the polyhedral bacteriophage G capsid. However, these strands are not visible toward the center of the zone of packaged DNA. 2) Within some positively stained particles, electron microscopy reveals DNA-associated stain in relatively high concentration at corners of the polyhedral bacteriophage G capsid. 3) When cross-linked DNA is expelled from its capsid during preparation for electron microscopy, some DNA molecules consist primarily of a compacted central region, surrounded by DNA strands that appear to be unravelling at multiple positions uniformly distributed around the compacted DNA region. The above results are explained by a previously presented model in which DNA is compacted by folding to form 12 icosahedrally arranged pear-shaped rings.
当双链DNA被包装在噬菌体衣壳中时,它占据一个腔,该腔的体积大约是DNA双螺旋体积的两倍。到目前为止,数据尚未揭示包装好的噬菌体DNA的紧密程度是通过DNA的折叠、DNA的单向缠绕,还是折叠和缠绕两者的结合来实现的。为了有助于区分这些可能性,本研究使用电子显微镜以及紫外线诱导的DNA-DNA交联,来获取关于包装在相对较大的噬菌体G中的DNA构象的以下信息:1)在噬菌体G的一些负染色颗粒的周边,电子显微镜显示出彼此平行且与多面体噬菌体G衣壳平行的DNA链。然而,在包装DNA区域的中心看不到这些链。2)在一些正染色颗粒内,电子显微镜显示在多面体噬菌体G衣壳的角处有相对高浓度的与DNA相关的染色。3)当在制备电子显微镜样品时将交联的DNA从其衣壳中排出时,一些DNA分子主要由一个紧密的中心区域组成,周围是DNA链,这些链似乎在围绕紧密DNA区域均匀分布的多个位置处解开。上述结果由先前提出的一个模型解释,在该模型中,DNA通过折叠被压缩以形成12个二十面体排列的梨形环。