Chattopadhyay Soma, Weller Sandra K
Department of Molecular, Microbial, and Structural Biology, MC3205, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA.
J Virol. 2007 Dec;81(24):13659-67. doi: 10.1128/JVI.01204-07. Epub 2007 Oct 17.
UL9, a superfamily II helicase, is a multifunctional protein required for herpes simplex virus type 1 replication in vivo. Although the C-terminal 317-amino-acid DNA binding domain of UL9 exists as a monomer, the full-length protein behaves as a dimer in solution. Thus, it has been assumed that the N-terminal 534 residues contain a region necessary for efficient dimerization and that UL9 dimers are in a head-to-head configuration. We recently showed, however, that residues in the N terminus could modulate the inhibitory properties of UL9 by decreasing the DNA binding ability of the C terminus (S. Chattopadhyay and S. K. Weller, J. Virol. 80:4491-4500, 2006). We suggested that a direct interaction between the N- and C-terminal portions of UL9 might exist and serve to modulate the DNA binding activities of the C terminus. In this study, we used a coimmunoprecipitation assay to show that the N-terminal portion of UL9 can indeed directly interact with the C terminus. A series of truncation mutant proteins were used to show that a region in the N terminus between residues 293 and 321 is necessary for efficient interaction. Similarly, a region in the C terminus between residues 600 and 800 is required for this interaction. The simplest model to explain these data is that UL9 dimers are oriented in a head-to-tail arrangement in which the N terminus is in contact with the C terminus.
UL9是一种超家族II解旋酶,是1型单纯疱疹病毒在体内复制所需的多功能蛋白。尽管UL9的C端317个氨基酸的DNA结合结构域以单体形式存在,但全长蛋白在溶液中表现为二聚体。因此,人们认为N端的534个残基包含有效二聚化所必需的区域,并且UL9二聚体呈头对头构型。然而,我们最近发现,N端的残基可以通过降低C端的DNA结合能力来调节UL9的抑制特性(S. Chattopadhyay和S. K. Weller,《病毒学杂志》80:4491 - 4500,2006)。我们推测UL9的N端和C端部分之间可能存在直接相互作用,并用于调节C端的DNA结合活性。在本研究中,我们使用共免疫沉淀试验表明UL9的N端部分确实可以与C端直接相互作用。一系列截短突变蛋白被用于表明N端293至321位残基之间的区域对于有效相互作用是必需 的。同样,C端600至800位残基之间的区域对于这种相互作用也是必需的。解释这些数据的最简单模型是,UL9二聚体以头对尾的排列方式定向,其中N端与C端接触。