Hang J Q, Catalano C E, Feiss M
Molecular Biology Program and Department of Microbiology, University of Iowa, Iowa City, Iowa 52242, USA.
Biochemistry. 2001 Nov 6;40(44):13370-7. doi: 10.1021/bi011126r.
cosN is the site at which terminase, the DNA packaging enzyme of phage lambda, introduces staggered nicks into viral concatemeric DNA to initiate genome packaging. Although the nick positions and many of the base pairs of cosN show 2-fold rotational symmetry, cosN is functionally asymmetric. That is, the cosN G2C mutation in the left half-site (cosNL) causes a strong virus growth defect whereas the symmetrically disposed cosN C11G mutation in the right half-site (cosNR) does not affect virus growth. The experiments reported here test the proposal that the genetic asymmetry of cosN results from terminase interactions with cosB, a binding site to the right of cosN. In the presence of cosB, the left half-site mutation, cosN G2C, strongly affected the cos cleavage reaction, while the symmetric right half-site mutation, cosN C11G, had little effect. In the absence of cosB, the two mutations moderately reduced the rate of cos cleavage by the same amount. The results indicated that the functional asymmetry of cosNdepends on the presence of cosB. A model is discussed in which terminase-cosN interactions in the nicking complex are assisted by anchoring of terminase to cosB.
cosN是噬菌体λ的DNA包装酶末端酶将交错切口引入病毒串联体DNA以启动基因组包装的位点。虽然cosN的切口位置和许多碱基对呈现2倍旋转对称,但cosN在功能上是不对称的。也就是说,左半位点(cosNL)的cosN G2C突变会导致强烈的病毒生长缺陷,而右半位点(cosNR)对称分布的cosN C11G突变则不影响病毒生长。本文报道的实验检验了以下提议,即cosN的遗传不对称性源于末端酶与cosB(cosN右侧的一个结合位点)的相互作用。在存在cosB的情况下,左半位点突变cosN G2C强烈影响cos切割反应,而对称的右半位点突变cosN C11G几乎没有影响。在不存在cosB的情况下,这两个突变以相同程度适度降低了cos切割速率。结果表明,cosN的功能不对称性取决于cosB的存在。文中讨论了一个模型,其中切口复合物中的末端酶-cosN相互作用通过末端酶与cosB的锚定得到辅助。