School of Biological Sciences, University of Auckland, Auckland, New Zealand.
Proc Natl Acad Sci U S A. 2009 Dec 29;106(52):22205-10. doi: 10.1073/pnas.0910686106. Epub 2009 Dec 10.
Baculoviruses are ubiquitous insect viruses well known for their use as bioinsecticides, gene therapy vectors, and protein expression systems. Overexpression of recombinant proteins in insect cell culture utilizes the strong promoter of the polyhedrin gene. In infected larvae, the polyhedrin protein forms robust intracellular crystals called polyhedra, which protect encased virions for prolonged periods in the environment. Polyhedra are produced by two unrelated families of insect viruses, baculoviruses and cypoviruses. The atomic structure of cypovirus polyhedra revealed an intricate packing of trimers, which are interconnected by a projecting N-terminal helical arm of the polyhedrin molecule. Baculovirus and cypovirus polyhedra share nearly identical lattices, and the N-terminal region of the otherwise unrelated baculovirus polyhedrin protein sequence is also predicted to be alpha-helical. These results suggest homology between the proteins and a common structural basis for viral polyhedra. Here, we present the 2.2-A structure of baculovirus polyhedra determined by x-ray crystallography from microcrystals produced in vivo. We show that the underlying molecular organization is, in fact, very different. Although both polyhedra have nearly identical unit cell dimensions and share I23 symmetry, the polyhedrin molecules are structurally unrelated and pack differently in the crystals. In particular, disulfide bonds and domain-swapped N-terminal domains stabilize the building blocks of baculovirus polyhedra and interlocking C-terminal arms join unit cells together. We show that the N-terminal projecting helical arms have different structural roles in baculovirus and cypovirus polyhedra and conclude that there is no structural evidence for a common evolutionary origin for both classes of polyhedra.
杆状病毒是一种普遍存在的昆虫病毒,以其作为生物杀虫剂、基因治疗载体和蛋白质表达系统而闻名。在昆虫细胞培养中,重组蛋白的过表达利用多角体蛋白基因的强启动子。在感染的幼虫中,多角体蛋白形成坚固的细胞内晶体,称为多角体,多角体在环境中长时间保护包裹的病毒粒子。多角体由两种不相关的昆虫病毒家族产生,杆状病毒和 cypovirus。cypovirus 多角体的原子结构揭示了三聚体的复杂包装,这些三聚体通过多角体分子的伸出的 N 端螺旋臂相互连接。杆状病毒和 cypovirus 多角体具有几乎相同的晶格,而不相关的杆状病毒多角体蛋白序列的 N 端区域也被预测为α-螺旋。这些结果表明蛋白质之间存在同源性和病毒多角体的共同结构基础。在这里,我们通过体内产生的微晶的 X 射线晶体学确定了杆状病毒多角体的 2.2-A 结构。我们表明,基础分子组织实际上非常不同。尽管两种多角体具有几乎相同的晶胞尺寸和共享 I23 对称性,但多角体蛋白在结构上是不相关的,并且在晶体中包装方式不同。特别是,二硫键和结构域交换的 N 端结构域稳定了杆状病毒多角体的构建块,并且交错的 C 端臂将单元细胞连接在一起。我们表明,N 端伸出的螺旋臂在杆状病毒和 cypovirus 多角体中具有不同的结构作用,并得出结论,没有结构证据表明这两类多角体具有共同的进化起源。