Edinburgh Explorer Functional analysis of the herpes simplex virus UL42 protein
Edinburgh Explorer Functional analysis of the herpes simplex virus UL42 protein
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单纯疱疹病毒UL42基因编码一种多功能多肽（UL42），对病毒DNA复制至关重要。为了进一步了解UL42的结构与其在病毒复制过程中所起的作用之间的关系，我们分析了一组广泛的突变UL42蛋白，以了解其执行三种主要生化功能的能力：与DNA结合，与病毒DNA聚合酶（Pol）稳定结合，以及增加Pol合成的DNA链的长度。选择的突变体也被检测其补充UL42无效病毒复制的能力。结果表明，UL42的n端340个氨基酸足以满足上述三种生化活性，并能支持病毒复制。进行性c端截断导致在Pol结合前失去可检测到的DNA结合活性，而多肽N端附近的几个突变导致与DNA的相互作用改变，但对Pol结合没有明显影响。更引人注目的是，残基160处的插入突变破坏了结合Pol的能力，但对DNA结合没有影响。这种改变的多肽也不能增加Pol合成的DNA产物的长度，并且突变基因不能补充UL42缺失病毒的生长，这表明Pol和ULA2之间的特异性相互作用对于Pol的完全功能和病毒复制是必要的。这项研究证实了Pol-UL42相互作用作为设计新型治疗剂的靶点的有效性。
The herpes simplex virus UL42 gene encodes a multifunctional polypeptide (UL42) that is essential for virus DNA replication. To further understand the relationship between the structure of UL42 and the role that it plays during virus replication, we analyzed an extensive set of mutant UL42 proteins for the ability to perform the three major biochemical functions ascribed to the protein: binding to DNA, stably associating with the virus DNA polymerase (Pol), and acting to increase the length of DNA chains synthesized by Pol. Selected mutants were also assayed for their ability to complement the replication of a UL42 null virus. The results indicated that the N-terminal 340 amino acids of UL42 were sufficient for all three biochemical activities and could also support virus replication. Progressive C-terminal truncation resulted in the loss of detectable DNA-binding activity before Pol binding, while several mutations near the N terminus of the polypeptide resulted in an altered interaction with DNA but had no apparent affect on Pol binding. More dramatically, an insertion mutation at residue 160 destroyed the ability to bind Pol but had no effect on DNA binding. This altered polypeptide also failed to increase the length of DNA product synthesized by Pol, and the mutant gene could not complement the growth of a UL42 null virus, indicating that the specific interaction between Pol and ULA2 is necessary for full Pol function and for virus replication. This study confirms the validity of the Pol-UL42 interaction as a target for the design of novel therapeutic agents.