Belyavskyi M, Braunagel S C, Summers M D
Department of Entomology, Texas A&M University, College Station, TX 77843-2475, USA.
Proc Natl Acad Sci U S A. 1998 Sep 15;95(19):11205-10. doi: 10.1073/pnas.95.19.11205.
Two major characteristics of baculovirus infection are arrest of the host cell at G2/M phase of the cell cycle with continuing viral DNA replication. We show that Autographa californica nucleopolyhedrovirus (AcMNPV) encodes for a multifunctional cyclin that may partially explain the molecular basis of these important characteristics of AcMNPV (baculovirus) infection. Amino acids 80-110 of the viral structural protein ODV-EC27 (-EC27) demonstrate 25-30% similarity with cellular cyclins within the cyclin box. Immunoprecipitation results using antibodies to -EC27 show that -EC27 can associate with either cdc2 or cdk6 resulting in active kinase complexes that can phosphorylate histone H1 and retinoblastoma protein in vitro. The cdk6-EC27 complex also associates with proliferating cell nuclear antigen (PCNA) and we demonstrate that PCNA is a structural protein of both the budded virus and the occlusion-derived virus. These results suggest that -EC27 can function as a multifunctional cyclin: when associated with cdc2, it exhibits cyclin B-like activity; when associated with cdk6, the complex possesses cyclin D-like activity and binds PCNA. The possible roles of such a multifunctional cyclin during the life cycle of baculovirus are discussed, along with potential implications relative to the expression of functionally authentic recombinant proteins by using baculovirus-infected cells.
杆状病毒感染的两个主要特征是宿主细胞在细胞周期的G2/M期停滞,同时病毒DNA持续复制。我们发现苜蓿银纹夜蛾核多角体病毒(AcMNPV)编码一种多功能细胞周期蛋白,这可能部分解释了AcMNPV(杆状病毒)感染这些重要特征的分子基础。病毒结构蛋白ODV-EC27(-EC27)的第80-110位氨基酸在细胞周期蛋白框内与细胞周期蛋白有25%-30%的相似性。使用针对-EC27的抗体进行免疫沉淀的结果表明,-EC27可与cdc2或cdk6结合,形成活性激酶复合物,该复合物在体外可磷酸化组蛋白H1和视网膜母细胞瘤蛋白。cdk6-EC27复合物还与增殖细胞核抗原(PCNA)结合,并且我们证明PCNA是出芽病毒和多角体衍生病毒的结构蛋白。这些结果表明,-EC27可作为一种多功能细胞周期蛋白发挥作用:当与cdc2结合时,它表现出类似细胞周期蛋白B的活性;当与cdk6结合时,该复合物具有类似细胞周期蛋白D的活性并结合PCNA。本文讨论了这种多功能细胞周期蛋白在杆状病毒生命周期中的可能作用,以及与利用杆状病毒感染细胞表达功能正确的重组蛋白的潜在意义。